Issue with selectively labelling genes in the Enhanced Volcano package
1
0
Entering edit mode
Jonathan ▴ 10
@6b06c9c3
Last seen 2.8 years ago
Germany

Hi all and thank you for your help in advance.

I am plotting the results of DESeq2 output using Enhanced Volcano package. I also want to plot only 20 gene labels.

My result table:

> head(resC3)
                 row  baseMean log2FoldChange    lfcSE        stat    pvalue      padj symbol                          geneName
1 ENSMUSG00000102693 1.5184259     -0.7800748 3.238902 -0.24084540 0.8096749 0.9920008   <NA>                              <NA>
2 ENSMUSG00000051951 4.4347206      0.7240531 1.907310  0.37962014 0.7042274 0.9896822   Xkr4 X-linked Kx blood group related 4
3 ENSMUSG00000102851 0.7409902     -3.2522220 3.440876 -0.94517258 0.3445708 0.9896822   <NA>                              <NA>
4 ENSMUSG00000103377 2.1128707      0.1741636 3.016754  0.05773211 0.9539620 0.9965102   <NA>                              <NA>
5 ENSMUSG00000104017 2.3202905      1.3844931 3.316122  0.41750370 0.6763100 0.9896822   <NA>                              <NA>
6 ENSMUSG00000103025 0.6405565      0.8586579 3.443174  0.24937975 0.8030670 0.9908850   <NA>                              <NA>

Rearrange the color:

keyvals.colour <- ifelse(
    resC3$log2FoldChange < 0 & resC3$padj < 0.1, 'royalblue',
      ifelse(resC3$log2FoldChange >= 0 & resC3$padj < 0.1, 'red2',
        'black'))
  keyvals.colour[is.na(keyvals.colour)] <- 'black'
  names(keyvals.colour)[keyvals.colour == 'red2'] <- 'Up-regulated'
  names(keyvals.colour)[keyvals.colour == 'black'] <- 'NS'
  names(keyvals.colour)[keyvals.colour == 'royalblue'] <- 'Down-regulated'

Select the labels to print:

selectLab<-resC3$symbol[names(keyvals.colour) %in% c('Up-regulated', 'Down-regulated')]
selectLab<-selectLab[!is.na(selectLab)]

str(selectLab)
chr [1:16] "Msc" "Knstrn" "Ovol2" "Olfml3" "Gstm2" "Lrrc17" "Oas1b" "Lrguk" "Tspan11" "Mrgpra3" "Plat" "Tmem184c" "Ppp2r1b" ...

Plot:

EC3<-EnhancedVolcano(resC3,
    subtitle = '',
    lab = resC3$symbol,
    x = 'log2FoldChange',
    y = 'padj',
    selectLab = selectLab,
    title = contrast,
    pCutoff = 0.0,
    FCcutoff = 0.0,
    pointSize = 2.0,
    labSize = 4.0,
    #col=c('grey30', 'grey30', 'royalblue', 'red2'),
    cutoffLineType = 'blank',
    cutoffLineCol = 'black',
    cutoffLineWidth = 0.8,
    hline = c(10e-2),
    hlineCol = c('black'),
    hlineType = c('longdash'),
    hlineWidth = c(0.6),
    vline = c(0),
    vlineCol = c('black'),
    vlineType = c('longdash'),
    vlineWidth = c(0.6),
    gridlines.major = FALSE,
    gridlines.minor = FALSE,
    colCustom = keyvals.colour) +
    theme(axis.text.y = element_text(size=10))

Only a few are plotted.

enter image description here

Could you help on this please? Many thanks.

sessionInfo( )

R version 4.0.0 (2020-04-24)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: CentOS Linux 7 (Core)

Matrix products: default
BLAS/LAPACK: /g/easybuild/x86_64/CentOS/7/haswell/software/OpenBLAS/0.3.9-GCC-9.3.0/lib/libopenblas_haswellp-r0.3.9.so

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C               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    LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C             LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] parallel  stats4    stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] ggbreak_0.0.8               biomaRt_2.44.1              DBI_1.1.0                   RSQLite_2.2.0              
 [5] GO.db_3.11.4                mgsa_1.36.0                 EnhancedVolcano_1.6.0       ggrepel_0.8.2              
 [9] knitr_1.28                  GGally_1.5.0                gridExtra_2.3               RColorBrewer_1.1-2         
[13] GenomicFeatures_1.40.1      scales_1.1.0                reshape2_1.4.4              org.Mm.eg.db_3.11.4        
[17] AnnotationDbi_1.50.1        pheatmap_1.0.12             gplots_3.0.3                genefilter_1.70.0          
[21] ggplot2_3.3.0               DESeq2_1.28.1               SummarizedExperiment_1.18.2 DelayedArray_0.14.0        
[25] matrixStats_0.56.0          Biobase_2.48.0              GenomicRanges_1.40.0        GenomeInfoDb_1.24.2        
[29] IRanges_2.22.2              S4Vectors_0.26.1            BiocGenerics_0.34.0        

loaded via a namespace (and not attached):
 [1] colorspace_1.4-1         ellipsis_0.3.0           XVector_0.28.0           aplot_0.1.1              rstudioapi_0.11         
 [6] farver_2.0.3             bit64_0.9-7              fansi_0.4.1              splines_4.0.0            cachem_1.0.6            
[11] geneplotter_1.66.0       Rsamtools_2.4.0          annotate_1.66.0          dbplyr_1.4.3             compiler_4.0.0          
[16] httr_1.4.2               assertthat_0.2.1         Matrix_1.3-4             fastmap_1.0.1            cli_2.0.2               
[21] htmltools_0.4.0          prettyunits_1.1.1        tools_4.0.0              gtable_0.3.0             glue_1.4.0              
[26] GenomeInfoDbData_1.2.3   dplyr_0.8.5              rappdirs_0.3.1           Rcpp_1.0.4.6             vctrs_0.2.4             
[31] Biostrings_2.56.0        gdata_2.18.0             rtracklayer_1.48.0       xfun_0.13                stringr_1.4.0           
[36] lifecycle_0.2.0          gtools_3.8.2             XML_3.99-0.3             zlibbioc_1.34.0          hms_0.5.3               
[41] yaml_2.2.1               curl_4.3                 memoise_2.0.0            ggfun_0.0.4              yulab.utils_0.0.4       
[46] reshape_0.8.8            stringi_1.4.6            highr_0.8                caTools_1.18.0           BiocParallel_1.22.0     
[51] rlang_0.4.5              pkgconfig_2.0.3          bitops_1.0-6             evaluate_0.14            lattice_0.20-41         
[56] purrr_0.3.4              GenomicAlignments_1.24.0 patchwork_1.0.0          htmlwidgets_1.5.1        labeling_0.3            
[61] bit_1.1-15.2             tidyselect_1.0.0         plyr_1.8.6               magrittr_1.5             R6_2.4.1                
[66] pillar_1.4.3             withr_2.2.0              survival_3.1-12          RCurl_1.98-1.2           tibble_3.0.1            
[71] crayon_1.3.4             KernSmooth_2.23-17       BiocFileCache_1.12.0     BisqueRNA_1.0.5          rmarkdown_2.11          
[76] progress_1.2.2           locfit_1.5-9.4           grid_4.0.0               data.table_1.12.8        blob_1.2.1              
[81] digest_0.6.25            xtable_1.8-4             gridGraphics_0.5-0       openssl_1.4.1            munsell_0.5.0           
[86] ggplotify_0.1.0          askpass_1.1
EnhancedVolcano • 1.9k views
ADD COMMENT
0
Entering edit mode
Kevin Blighe ★ 4.0k
@kevin
Last seen 9 days ago
Republic of Ireland

Hi, what is the output of:

str(resC3)
length(selectLab)
which(selectLab %in% resC3$symbol)

By the way, you may want to plot un-adjusted p-values, not adjusted ones.

ADD COMMENT
0
Entering edit mode

Hi Kevin, Thanks for your reply. Here ate the requested output:

> str(resC3)
'data.frame':   26154 obs. of  9 variables:
 $ row           : chr  "ENSMUSG00000102693" "ENSMUSG00000051951" "ENSMUSG00000102851" "ENSMUSG00000103377" ...
 $ baseMean      : num  1.518 4.435 0.741 2.113 2.32 ...
 $ log2FoldChange: num  -0.78 0.724 -3.252 0.174 1.384 ...
 $ lfcSE         : num  3.24 1.91 3.44 3.02 3.32 ...
 $ stat          : num  -0.2408 0.3796 -0.9452 0.0577 0.4175 ...
 $ pvalue        : num  0.81 0.704 0.345 0.954 0.676 ...
 $ padj          : num  0.992 0.99 0.99 0.997 0.99 ...
 $ symbol        : chr  NA "Xkr4" NA NA ...
 $ geneName      : chr  NA "X-linked Kx blood group related 4" NA NA ...
> length(selectLab)
[1] 16
> which(selectLab %in% resC3$symbol)
 [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16

Thanks for your help.

ADD REPLY
0
Entering edit mode

It can be that there is no space to plot all labels. Can you try drawConnectors = TRUE

ADD REPLY
1
Entering edit mode

It does the trick, thanks!

enter image description here

ADD REPLY

Login before adding your answer.

Traffic: 823 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6