Last updated: 2024-10-12

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Knit directory: GibbsDTE-code/analysis/

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Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes.


These are the previous versions of the repository in which changes were made to the R Markdown (analysis/simulation-supp.Rmd) and HTML (docs/simulation-supp.html) files. If you’ve configured a remote Git repository (see ?wflow_git_remote), click on the hyperlinks in the table below to view the files as they were in that past version.

File Version Author Date Message
Rmd d1585c4 Pedro Baldoni 2024-10-12 Updating Supp Figure S9 to the same format as in the main text
html d7807a8 Pedro Baldoni 2024-10-09 Build site.
Rmd 529139f Pedro Baldoni 2024-10-09 NARGAB Revision from October 2024
html 0f05c72 Pedro Baldoni 2024-06-24 Build site.
Rmd 5b41ed5 Pedro Baldoni 2024-06-24 Printing out tables
html 70ab63f Pedro Baldoni 2024-04-05 Publishing workflowr
Rmd bebf946 Pedro Baldoni 2024-04-05 Adding simulation Rmds

Introduction

This Rmarkdown generates Supplementary Figures S4–S9 and Supplementary Table S1 from our Gibbs DTE paper. The results are based on the simulation designed to emulate small-scale experiments with low biological variation and sample size, a typical scenario of RNA-seq studies with genetically modified organisms or cell lines.

Setup

Below we set options for this Rmarkdown report and load the necessary libraries.

knitr::opts_chunk$set(dev = "png",
                      dpi = 300,
                      dev.args = list(type = "cairo-png"),
                      root.dir = '.',
                      autodep = TRUE)

options(knitr.kable.NA = "-")

bs <- 8
library(data.table)
library(magrittr)
library(ggplot2)
library(patchwork)
library(plyr)
library(kableExtra)
devtools::load_all('../code/pkg')

Next we set the paths to our simulation results.

path.misc <- file.path('../misc/simulation-supp.Rmd')
dir.create(path.misc,recursive = TRUE,showWarnings = FALSE)

path.example <- "../output/simulation/data/mm39/readlen-100/fc2/paired-end/9999TxPerGene/unbalanced/5libsPerGroup"

path.fdr <-
  list.files('../output/simulation/summary','fdr.tsv.gz',recursive = TRUE,full.names = TRUE)
path.metrics <-
  list.files('../output/simulation/summary','metrics.tsv.gz',recursive = TRUE,full.names = TRUE)
path.time <-
  list.files('../output/simulation/summary','^time.tsv.gz',recursive = TRUE,full.names = TRUE)
path.quanttime <-
  list.files('../output/simulation/summary','quanttime.tsv.gz',recursive = TRUE,full.names = TRUE)
path.quantile <-
  list.files('../output/simulation/summary','quantile.tsv.gz',recursive = TRUE,full.names = TRUE)
path.pvalue <-
  list.files('../output/simulation/summary','pvalue.tsv.gz',recursive = TRUE,full.names = TRUE)
path.overdispersion <-
  list.files('../output/simulation/summary','overdispersion.tsv.gz',recursive = TRUE,full.names = TRUE)
path.roc <-
  list.files('../output/simulation/summary','roc.tsv.gz',recursive = TRUE,full.names = TRUE)

Data loading

Below we load the actual results and relabel some of the factor variables.

relabel <- function(x){
  y <- copy(x)
  y$TxPerGene %<>% factor(levels = paste0(c(2, 3, 4, 5, 9999), 'TxPerGene'),labels = c(paste0("#Tx/Gene = ", c(2, 3, 4, 5)), 'All Transcripts')) %<>% droplevels()
  y$LibsPerGroup %<>% factor(levels = paste0(c(3, 5, 10), 'libsPerGroup'),labels = paste0('#Lib/Group = ', c(3, 5, 10))) %<>% droplevels()
  y$Quantifier %<>% factor(levels = c('salmon','salmon-gibbs','kallisto'),labels = c('Salmon-Boot','Salmon-Gibbs','kallisto')) %<>% droplevels()
  y$Length %<>% factor(levels = paste0('readlen-', seq(50, 150, 25)),labels = paste0(seq(50, 150, 25), 'bp')) %<>% droplevels()
  if (!is.null(x$Sample)) {
    y$Sample %<>% factor(levels = c(paste0("groupA_rep",1:10,"_R1"),paste0("groupB_rep",1:10,"_R1")),labels = c(paste0("A",1:10),paste0("B",1:10))) %<>% droplevels()
  }
  return(y)
}

# Loading datasets
dt.fdr <- do.call(rbind,lapply(path.fdr,fread)) %>% relabel()
dt.metrics <- do.call(rbind,lapply(path.metrics,fread)) %>% relabel()
dt.time <- do.call(rbind,lapply(path.time,fread)) %>% relabel()
dt.quanttime <- do.call(rbind,lapply(path.quanttime,fread)) %>% relabel()
dt.quantile <- do.call(rbind,lapply(path.quantile,fread)) %>% relabel()
dt.pvalue <- do.call(rbind,lapply(path.pvalue,fread)) %>% relabel()
dt.overdispersion <- do.call(rbind,lapply(path.overdispersion,fread)) %>% relabel()
dt.roc <- do.call(rbind,lapply(path.roc,fread)) %>% relabel()

Power

Below I compute the barplots of Supplementary Figures S4–S8.

dt.power <- dt.metrics[FC == 'fc2' &
                         !grepl("\\.n|RC",Method) &
                         Scenario == 'unbalanced',]

dt.power$LibsPerGroup %<>% factor(levels = paste0('#Lib/Group = ', c(3, 5,10)),labels = paste0(c(3,5,10),' samples per group'))
dt.power$Scenario %<>% factor(levels = c('balanced','unbalanced'),labels = c('Equal library sizes','Unequal library sizes'))
dt.power$Quantifier %<>% 
  factor(levels = c('kallisto','Salmon-Boot','Salmon-Gibbs'),
         labels = c('Bootstrap\nkallisto','Bootstrap\nSalmon','Gibbs\nSalmon'))

dt.power$Method %<>% mapvalues(from = c('edgeR-SC','edgeR.legacy-SC'),to = c('edgeR-v4','edgeR-v3'))

dt.power[, FDR := roundPretty(ifelse((FP+TP) == 0,NA,100*FP/(FP+TP)),1)]

plot.power <- function(df.bar,df.txt,scenario,library,legend = FALSE, base_size = bs,...){
  tb.bar <- df.bar[Scenario == scenario & LibsPerGroup == library,]
  tb.txt <- df.txt[Scenario == scenario & LibsPerGroup == library,][FDR != 'NA',]
  gap <- 0.05*max(dt.power$TP + dt.power$FP)
  ggplot(tb.bar,aes(x = Quantifier,y = Value,fill = Type)) +
    facet_wrap(facets = "Method",nrow = 1) +
    geom_col() +
    geom_text(aes(x = Quantifier,y = (TP + FP) +  gap,label = FDR),
              vjust = 0,data = tb.txt,size = base_size/.pt,inherit.aes = FALSE) +
    scale_fill_manual(values = c('#ff0000','#bebebe')) +
    labs(x = NULL,y = paste('DE Transcripts')) +
    coord_cartesian(...) +
    theme_bw(base_size = base_size,base_family = 'sans') +
    theme(panel.grid = element_blank(),
          # axis.text.x = element_text(angle = 90),
          axis.text = element_text(colour = 'black',size = base_size),
          strip.text = element_text(size = base_size)) +
    if (legend == TRUE) theme(legend.background = element_rect(fill = alpha('white', 0)),
                              legend.text = element_text(size = base_size),
                              legend.position = 'inside',
                              legend.position.inside = c(0.95,0.825),
                              legend.title = element_blank(),
                              legend.key.size = unit(0.75,"line")) else theme(legend.position = 'none')
}

make.plot.power <- function(x,scenario,library,quantifier,...){
  x.melt <- melt(x,id.vars = colnames(x)[-which(colnames(x) %in% c('P.SIG','TP','FP'))],
                 measure.vars = c('TP','FP'),
                 variable.name = 'Type',
                 value.name = 'Value')
  
  x.melt <- x.melt[Quantifier %in% quantifier,]
  y <- copy(x[Quantifier %in% quantifier,])
  
  x.melt$Quantifier <- droplevels(x.melt$Quantifier)
  y$Quantifier <- droplevels(y$Quantifier)
  
  x.melt$Type <-
    factor(x.melt$Type,
           levels = c('FP','TP'),
           labels = c('False','True'))
  
  plot.power(df.bar = x.melt,
             df.txt = y,
             scenario = scenario,
             library = library,
             legend = TRUE,
             ...)
}


fig.power.salmon.3 <- make.plot.power(x = dt.power,
                                      scenario = 'Unequal library sizes',
                                      library = '3 samples per group',
                                      quantifier = c('Bootstrap\nSalmon',
                                                     'Gibbs\nSalmon'),
                                      ylim = c(0,3250))
fig.power.salmon.10 <- make.plot.power(x = dt.power,
                                       scenario = 'Unequal library sizes',
                                       library = '10 samples per group',
                                       quantifier = c('Bootstrap\nSalmon',
                                                      'Gibbs\nSalmon'),
                                       ylim = c(0,5000))

fig.power.kallisto.3 <- make.plot.power(x = dt.power,
                                        scenario = 'Unequal library sizes',
                                        library = '3 samples per group',
                                        quantifier = c('Bootstrap\nkallisto',
                                                       'Gibbs\nSalmon'),
                                        ylim = c(0,3250))
fig.power.kallisto.5 <- make.plot.power(x = dt.power,
                                        scenario = 'Unequal library sizes',
                                        library = '5 samples per group',
                                        quantifier = c('Bootstrap\nkallisto',
                                                       'Gibbs\nSalmon'),
                                        ylim = c(0,3250))
fig.power.kallisto.10 <- make.plot.power(x = dt.power,
                                         scenario = 'Unequal library sizes',
                                         library = '10 samples per group',
                                         quantifier = c('Bootstrap\nkallisto',
                                                        'Gibbs\nSalmon'),
                                         ylim = c(0,5000))

fig.power.salmon.3

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.power.salmon.10

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.power.kallisto.3

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.power.kallisto.5

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.power.kallisto.10

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05

FDR

Below I compute the false discovery curves of Supplementary Figures S4–S8.

dt.fdr.plot <- dt.fdr[FC == 'fc2' &
                        !grepl("\\.n|RC",Method) &
                        Scenario == 'unbalanced',]

colmeth <- methodsNames()$color
names(colmeth) <- gsub('^edgeR-SC$','edgeR-v4',names(colmeth))
names(colmeth) <- gsub('^edgeR.legacy-SC$','edgeR-v3',names(colmeth))

dt.fdr.plot$LibsPerGroup %<>% factor(levels = paste0('#Lib/Group = ', c(3, 5,10)),labels = paste0(c(3,5,10),' samples per group'))
dt.fdr.plot$Scenario %<>% factor(levels = c('balanced','unbalanced'),labels = c('Equal library sizes','Unequal library sizes'))
dt.fdr.plot$Quantifier %<>% 
  factor(levels = c('Salmon-Gibbs','kallisto','Salmon-Boot'),
         labels = c('Gibbs\nSalmon','Bootstrap\nkallisto','Bootstrap\nSalmon'))
dt.fdr.plot$Method %<>% mapvalues(from = c('edgeR-SC','edgeR.legacy-SC'),to = c('edgeR-v4','edgeR-v3'))

dt.fdr.plot$Method %<>% factor(levels = c('Swish',
                                          'sleuth-Wald',
                                          'sleuth-LRT',
                                          'edgeR-v3',
                                          'edgeR-v4'))

plot.fdr <- function(df.line,scenario,library,legend = FALSE,base_size = bs,color = colmeth,...){
  tb.bar <- df.line[Scenario == scenario & LibsPerGroup == library,]
  
  ggplot(tb.bar,aes(x = N,y = FDR,color = Method,linetype = Quantifier)) +
    geom_line(linewidth = 0.7) +
    scale_color_manual(values = color,
                       breaks = c('edgeR-v3',
                                  'edgeR-v4',
                                  'sleuth-LRT',
                                  'sleuth-Wald',
                                  'Swish')) +
    coord_cartesian(...) +
    labs(y = 'False discoveries',x = 'Transcripts chosen') +
    theme_bw(base_size = base_size,base_family = 'sans') +
    theme(panel.grid = element_blank(),
          axis.text = element_text(colour = 'black',size = base_size)) +
    if (legend == TRUE) theme(legend.background = element_rect(fill = alpha('white', 0)),
                              legend.direction = 'vertical',
                              legend.box = 'horizontal',
                              legend.position = c(0.2,0.9),
                              legend.text = element_text(size = base_size),
                              legend.title = element_blank(),
                              legend.key.size = unit(0.75,"line")) else theme(legend.position = 'none')
}

fig.fdr.salmon.3 <- plot.fdr(df.line = dt.fdr.plot[Quantifier %in% c('Bootstrap\nSalmon','Gibbs\nSalmon'),],
                             scenario = 'Unequal library sizes',
                             library = '3 samples per group',
                             ylim = c(0,750),
                             xlim = c(0,2500),
                             legend = TRUE)
Warning: A numeric `legend.position` argument in `theme()` was deprecated in ggplot2
3.5.0.
ℹ Please use the `legend.position.inside` argument of `theme()` instead.
This warning is displayed once every 8 hours.
Call `lifecycle::last_lifecycle_warnings()` to see where this warning was
generated.
fig.fdr.salmon.10 <- plot.fdr(df.line = dt.fdr.plot[Quantifier %in% c('Bootstrap\nSalmon','Gibbs\nSalmon'),],
                              scenario = 'Unequal library sizes',
                              library = '10 samples per group',
                              ylim = c(0,300),
                              xlim = c(1000,3000),
                              legend = TRUE)

fig.fdr.kallisto.3 <- plot.fdr(df.line = dt.fdr.plot[Quantifier %in% c('Bootstrap\nkallisto','Gibbs\nSalmon'),],
                               scenario = 'Unequal library sizes',
                               library = '3 samples per group',
                               ylim = c(0,750),
                               xlim = c(0,2500),
                               legend = TRUE)

fig.fdr.kallisto.5 <- plot.fdr(df.line = dt.fdr.plot[Quantifier %in% c('Bootstrap\nkallisto','Gibbs\nSalmon'),],
                               scenario = 'Unequal library sizes',
                               library = '5 samples per group',
                               ylim = c(0,400),
                               xlim = c(0,2500),
                               legend = TRUE)

fig.fdr.kallisto.10 <- plot.fdr(df.line = dt.fdr.plot[Quantifier %in% c('Bootstrap\nkallisto','Gibbs\nSalmon'),],
                                scenario = 'Unequal library sizes',
                                library = '10 samples per group',
                                ylim = c(0,300),
                                xlim = c(1000,3000),
                                legend = TRUE)

fig.fdr.salmon.3

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.fdr.salmon.10

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.fdr.kallisto.3

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.fdr.kallisto.5

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05
fig.fdr.kallisto.10

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05

ROC curves

Below is the code chunk used to generate Supplementary Figure S9.

dt.rcurve <- dt.roc[FC == 'fc2' &
                      grepl("edgeR-SC|edgeR-SC\\.n",Method) &
                      Scenario == 'unbalanced',]

dt.rcurve$LibsPerGroup %<>% factor(levels = paste0('#Lib/Group = ', c(3, 5, 10)),labels = paste0(c('Three','Five', 'Ten'),' samples per group'))
dt.rcurve$Scenario %<>% factor(levels = c('balanced','unbalanced'),labels = c('Equal library sizes','Unequal library sizes'))
dt.rcurve$Quantifier %<>% factor(levels = c('kallisto','Salmon-Boot','Salmon-Gibbs'),labels = c('Bootstrap (kallisto)','Bootstrap (Salmon)','Gibbs (Salmon)'))
dt.rcurve$Method %<>%
  factor(levels = c(paste0('edgeR-SC.n',seq(10,90,10)),'edgeR-SC')) %<>%
  mapvalues(from = c(paste0('edgeR-SC.n',seq(10,90,10)),'edgeR-SC'),to = paste0(seq(10,100,10)))

dt.rcurve.long <- melt(dt.rcurve,
                       id.vars = c('Genome','Length','FC','Reads','TxPerGene','Scenario','LibsPerGroup','Quantifier','Method','nFDR'),
                       measure.vars = c('oTPR','oFDR'),
                       variable.name = 'Type',
                       value.name = 'Value')

foo <- function(x,lib,type,ylab,title = NULL,base_size = bs,legend = TRUE,...){
  sub.x <- x[LibsPerGroup == lib & Type == type,]
  
  sub.x$nFDR_char <- roundPretty(sub.x$nFDR,digits = 2)
  sub.x$nFDR_char %<>% factor(levels = c('0.01','0.05','0.10','0.15','0.20'))
  
  shape <- 21:25
  names(shape) <- c('0.01','0.05','0.10','0.15','0.20')
  
  p <- ggplot(data = sub.x, aes(x = Method,y = Value,group = interaction(nFDR_char,Quantifier))) +
    geom_line(aes(color = Quantifier),linewidth = 0.25) +
    geom_point(aes(shape = nFDR_char,color = Quantifier,fill = Quantifier),size = 1.25) +
    theme_bw(base_size = bs,base_family = 'sans') +
    scale_color_manual(values = c('Bootstrap (kallisto)' = 'green3','Bootstrap (Salmon)' = 'red','Gibbs (Salmon)' = 'blue')) +
    scale_fill_manual(values = c('Bootstrap (kallisto)' = 'green3','Bootstrap (Salmon)' = 'red','Gibbs (Salmon)' = 'blue'),guide = "none") +
    scale_shape_manual(values = shape) +
    coord_cartesian(...) +
    theme(panel.grid = element_blank(),
          plot.title = element_text(hjust = 0.5,size = base_size),
          axis.text = element_text(colour = 'black',size = base_size),
          strip.text = element_text(colour = 'black',size = base_size)) +
    labs(x = 'Number of Resamples',y = ylab,shape = 'Nominal FDR',color = 'Method',title = title) +
    if (legend == TRUE) {theme(legend.background = element_rect(fill = alpha('white', 0)),
                               legend.text = element_text(size = base_size),
                               legend.title = element_text(size = base_size),
                               legend.margin = margin(0, 0, 0, 0))} else theme(legend.position = 'none')
  p
}

fig.roc.three.tpr <- foo(dt.rcurve.long,lib = 'Three samples per group',type = 'oTPR',ylab = 'Observed Power',ylim = c(0.1,0.7)) + labs(x = NULL,title = "n = 3")
fig.roc.five.tpr <- foo(dt.rcurve.long,lib = 'Five samples per group',type = 'oTPR',ylab = 'Observed Power',ylim = c(0.55,0.9)) + labs(x = NULL,y = NULL,title = "n = 5")
fig.roc.ten.tpr <- foo(dt.rcurve.long,lib = 'Ten samples per group',type = 'oTPR',ylab = 'Observed Power',ylim = c(0.875,0.975)) + labs(x = NULL,y = NULL,title = "n = 10") + scale_y_continuous(breaks = c(0.88,0.92,0.96))

fig.roc.three.fdr <- foo(dt.rcurve.long,lib = 'Three samples per group',type = 'oFDR',ylab = 'Observed FDR',ylim = c(0,0.2))
fig.roc.five.fdr <- foo(dt.rcurve.long,lib = 'Five samples per group',type = 'oFDR',ylab = 'Observed FDR',ylim = c(0,0.2)) + labs(y = NULL)
fig.roc.ten.fdr <- foo(dt.rcurve.long,lib = 'Ten samples per group',type = 'oFDR',ylab = 'Observed FDR',ylim = c(0,0.2)) + labs(y = NULL)

wrap_plots(A = fig.roc.three.tpr,
           B = fig.roc.five.tpr,
           C = fig.roc.ten.tpr,
           D = fig.roc.three.fdr,
           E = fig.roc.five.fdr,
           `F` = fig.roc.ten.fdr,
           design = c(area(1,1),area(1,2),area(1,3),
                      area(2,1),area(2,2),area(2,3))) + 
  plot_layout(guides = 'collect') & theme(
    legend.position = 'bottom',
    legend.direction = 'horizontal',
    legend.box = 'vertical')

Version Author Date
d7807a8 Pedro Baldoni 2024-10-09
70ab63f Pedro Baldoni 2024-04-05

Quantification time

Below I generate Supplementary Table S1.

dt.salmontime <- dt.quanttime[,
                              .(TotalTime = mean(Time)/60,
                                ResamplingTime = mean(ResamplingTime)/60),
                              by = c('LibSize','Quantifier')]

dt.salmontime$QuantTime <- dt.salmontime$TotalTime - dt.salmontime$ResamplingTime
setcolorder(dt.salmontime,c('Quantifier','LibSize','QuantTime','ResamplingTime','TotalTime'))

dt.salmontime$LibSize %<>% factor(levels = c('2.5e+07','5e+07','1e+08'),
                                  labels = c('25','50','100'))

dt.salmontime$Quantifier %<>% factor(levels = c('kallisto','Salmon-Boot','Salmon-Gibbs'),
                                     labels = c('Bootstrap (kallisto)','Bootstrap (Salmon)','Gibbs (Salmon)'))

dt.salmontime <- dt.salmontime[order(Quantifier,LibSize),]

cnames <- c('Resampling\nMethod',
            'Library Size\n(million read pairs)',
            'Optimization',
            'Resampling',
            'Total')

tb.salmontime <- kbl(x = dt.salmontime,
                     format = 'latex',
                     digits = 2,
                     col.names = linebreak(cnames,align = 'c'),
                     align = 'ccccc',
                     escape = FALSE,
                     booktabs = TRUE) %>%
  add_header_above(c(" " = 2, "Quantification Time (minutes)" = 3)) %>%
  collapse_rows(1, latex_hline = 'major')

kbl(x = dt.salmontime,
    format = 'pipe',
    digits = 2,
    col.names = linebreak(cnames,align = 'c'),
    align = 'ccccc',
    escape = FALSE,
    booktabs = TRUE) %>%
  add_header_above(c(" " = 2, "Quantification Time (minutes)" = 3)) %>%
  collapse_rows(1, latex_hline = 'major')
Quantification Time (minutes)
Optimization Resampling Total
Bootstrap (kallisto) 25
4.92
50
7.32
100
11.71
Bootstrap (Salmon) 25 1.79 9.45 11.24
50 3.22 13.41 16.64
100 6.09 19.15 25.24
Gibbs (Salmon) 25 1.78 0.91 2.69
50 3.22 1.62 4.85
100 6.05 3.01 9.06

Output files

And finally all files are exported to be incorporated in the supplement.

fig.metrics.salmon.3 <- wrap_plots(A = fig.power.salmon.3,
                                   B = fig.fdr.salmon.3,
                                   design = c(area(1,1),area(2,1)),
                                   heights = c(0.25,0.75)) + 
  plot_annotation(tag_levels = 'a',
                  theme = theme(plot.tag = element_text(size = bs)))

fig.metrics.salmon.10 <- wrap_plots(A = fig.power.salmon.10,
                                    B = fig.fdr.salmon.10,
                                    design = c(area(1,1),area(2,1)),
                                    heights = c(0.25,0.75)) + 
  plot_annotation(tag_levels = 'a',
                  theme = theme(plot.tag = element_text(size = bs)))

fig.metrics.kallisto.3 <- wrap_plots(A = fig.power.kallisto.3,
                                     B = fig.fdr.kallisto.3,
                                     design = c(area(1,1),area(2,1)),
                                     heights = c(0.25,0.75)) + 
  plot_annotation(tag_levels = 'a',
                  theme = theme(plot.tag = element_text(size = bs)))

fig.metrics.kallisto.5 <- wrap_plots(A = fig.power.kallisto.5,
                                     B = fig.fdr.kallisto.5,
                                     design = c(area(1,1),area(2,1)),
                                     heights = c(0.25,0.75)) + 
  plot_annotation(tag_levels = 'a',
                  theme = theme(plot.tag = element_text(size = bs)))

fig.metrics.kallisto.10 <- wrap_plots(A = fig.power.kallisto.10,
                                      B = fig.fdr.kallisto.10,
                                      design = c(area(1,1),area(2,1)),
                                      heights = c(0.25,0.75)) + 
  plot_annotation(tag_levels = 'a',
                  theme = theme(plot.tag = element_text(size = bs)))

ggsave(plot = fig.metrics.salmon.3,filename = file.path(path.misc,'SuppFigure-Metrics-Salmon-3.pdf'),
       device = 'pdf',width = 6,height = 8,units = 'in',dpi = 300)
ggsave(plot = fig.metrics.salmon.10,filename = file.path(path.misc,'SuppFigure-Metrics-Salmon-10.pdf'),
       device = 'pdf',width = 6,height = 8,units = 'in',dpi = 300)

ggsave(plot = fig.metrics.kallisto.3,filename = file.path(path.misc,'SuppFigure-Metrics-kallisto-3.pdf'),
       device = 'pdf',width = 6,height = 8,units = 'in',dpi = 300)
ggsave(plot = fig.metrics.kallisto.5,filename = file.path(path.misc,'SuppFigure-Metrics-kallisto-5.pdf'),
       device = 'pdf',width = 6,height = 8,units = 'in',dpi = 300)
ggsave(plot = fig.metrics.kallisto.10,filename = file.path(path.misc,'SuppFigure-Metrics-kallisto-10.pdf'),
       device = 'pdf',width = 6,height = 8,units = 'in',dpi = 300)

fig.roc <- wrap_plots(A = fig.roc.three.tpr,
           B = fig.roc.five.tpr,
           C = fig.roc.ten.tpr,
           D = fig.roc.three.fdr,
           E = fig.roc.five.fdr,
           `F` = fig.roc.ten.fdr,
           design = c(area(1,1),area(1,2),area(1,3),
                      area(2,1),area(2,2),area(2,3))) + 
  plot_layout(guides = 'collect') & theme(
    legend.position = 'bottom',
    legend.direction = 'horizontal',
    legend.box = 'vertical')

ggsave(plot = fig.roc,filename = file.path(path.misc,'SuppFigure-ROC.pdf'),
       device = 'pdf',width = 7,height = 5,units = 'in',dpi = 300)

save_kable(tb.salmontime,file = file.path(path.misc,'SuppTable-QuantTime.tex'))

sessionInfo()
R version 4.3.3 (2024-02-29)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Red Hat Enterprise Linux 9.4 (Plow)

Matrix products: default
BLAS:   /stornext/System/data/software/rhel/9/base/tools/R/4.3.3/lib64/R/lib/libRblas.so 
LAPACK: /stornext/System/data/software/rhel/9/base/tools/R/4.3.3/lib64/R/lib/libRlapack.so;  LAPACK version 3.11.0

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

time zone: Australia/Melbourne
tzcode source: system (glibc)

attached base packages:
[1] stats     graphics  grDevices datasets  utils     methods   base     

other attached packages:
[1] pkg_1.0           kableExtra_1.4.0  plyr_1.8.9        patchwork_1.3.0  
[5] ggplot2_3.5.1     magrittr_2.0.3    data.table_1.16.0 workflowr_1.7.1  

loaded via a namespace (and not attached):
  [1] splines_4.3.3                 later_1.3.2                  
  [3] BiocIO_1.12.0                 bitops_1.0-8                 
  [5] filelock_1.0.3                R.oo_1.26.0                  
  [7] tibble_3.2.1                  XML_3.99-0.17                
  [9] lifecycle_1.0.4               edgeR_4.0.16                 
 [11] rprojroot_2.0.4               processx_3.8.4               
 [13] lattice_0.22-5                ensembldb_2.26.0             
 [15] limma_3.58.1                  sass_0.4.9                   
 [17] rmarkdown_2.28                jquerylib_0.1.4              
 [19] yaml_2.3.10                   remotes_2.5.0                
 [21] httpuv_1.6.15                 sessioninfo_1.2.2            
 [23] pkgbuild_1.4.4                DBI_1.2.3                    
 [25] abind_1.4-8                   pkgload_1.4.0                
 [27] zlibbioc_1.48.2               GenomicRanges_1.54.1         
 [29] R.utils_2.12.3                purrr_1.0.2                  
 [31] AnnotationFilter_1.26.0       BiocGenerics_0.48.1          
 [33] RCurl_1.98-1.16               rappdirs_0.3.3               
 [35] git2r_0.33.0                  GenomeInfoDbData_1.2.11      
 [37] wasabi_1.0.1                  IRanges_2.36.0               
 [39] S4Vectors_0.40.2              fishpond_2.8.0               
 [41] svglite_2.1.3                 codetools_0.2-19             
 [43] DelayedArray_0.28.0           xml2_1.3.6                   
 [45] tidyselect_1.2.1              farver_2.1.2                 
 [47] matrixStats_1.4.1             stats4_4.3.3                 
 [49] BiocFileCache_2.10.2          GenomicAlignments_1.38.2     
 [51] jsonlite_1.8.9                ellipsis_0.3.2               
 [53] systemfonts_1.1.0             tools_4.3.3                  
 [55] progress_1.2.3                ragg_1.3.3                   
 [57] Rcpp_1.0.13                   glue_1.8.0                   
 [59] svMisc_1.2.3                  SparseArray_1.2.4            
 [61] xfun_0.47                     MatrixGenerics_1.14.0        
 [63] usethis_3.0.0                 GenomeInfoDb_1.38.8          
 [65] dplyr_1.1.4                   withr_3.0.1                  
 [67] BiocManager_1.30.25           fastmap_1.2.0                
 [69] rhdf5filters_1.14.1           fansi_1.0.6                  
 [71] callr_3.7.6                   digest_0.6.37                
 [73] R6_2.5.1                      mime_0.12                    
 [75] textshaping_0.4.0             colorspace_2.1-1             
 [77] gtools_3.9.5                  biomaRt_2.58.2               
 [79] RSQLite_2.3.7                 R.methodsS3_1.8.2            
 [81] utf8_1.2.4                    generics_0.1.3               
 [83] renv_1.0.9                    tximeta_1.20.3               
 [85] rtracklayer_1.62.0            prettyunits_1.2.0            
 [87] httr_1.4.7                    htmlwidgets_1.6.4            
 [89] S4Arrays_1.2.1                whisker_0.4.1                
 [91] pkgconfig_2.0.3               gtable_0.3.5                 
 [93] blob_1.2.4                    SingleCellExperiment_1.24.0  
 [95] XVector_0.42.0                htmltools_0.5.8.1            
 [97] profvis_0.4.0                 ProtGenerics_1.34.0          
 [99] sleuth_0.30.1                 scales_1.3.0                 
[101] Biobase_2.62.0                Rsubread_2.16.1              
[103] png_0.1-8                     knitr_1.48                   
[105] rstudioapi_0.16.0             tzdb_0.4.0                   
[107] rjson_0.2.23                  curl_5.2.3                   
[109] cachem_1.1.0                  rhdf5_2.46.1                 
[111] stringr_1.5.1                 BiocVersion_3.18.1           
[113] parallel_4.3.3                miniUI_0.1.1.1               
[115] AnnotationDbi_1.64.1          restfulr_0.0.15              
[117] desc_1.4.3                    pillar_1.9.0                 
[119] grid_4.3.3                    vctrs_0.6.5                  
[121] urlchecker_1.0.1              promises_1.3.0               
[123] dbplyr_2.5.0                  xtable_1.8-4                 
[125] tximport_1.30.0               evaluate_1.0.0               
[127] readr_2.1.5                   GenomicFeatures_1.54.4       
[129] locfit_1.5-9.10               cli_3.6.3                    
[131] compiler_4.3.3                Rsamtools_2.18.0             
[133] rlang_1.1.4                   crayon_1.5.3                 
[135] labeling_0.4.3                ps_1.8.0                     
[137] getPass_0.2-4                 fs_1.6.4                     
[139] stringi_1.8.4                 viridisLite_0.4.2            
[141] BiocParallel_1.36.0           munsell_0.5.1                
[143] Biostrings_2.70.3             lazyeval_0.2.2               
[145] devtools_2.4.5                Matrix_1.6-5                 
[147] hms_1.1.3                     bit64_4.5.2                  
[149] Rhdf5lib_1.24.2               KEGGREST_1.42.0              
[151] statmod_1.5.0                 shiny_1.9.1                  
[153] highr_0.11                    SummarizedExperiment_1.32.0  
[155] interactiveDisplayBase_1.40.0 AnnotationHub_3.10.1         
[157] memoise_2.0.1                 thematic_0.1.6               
[159] bslib_0.8.0                   bit_4.5.0