diff --git a/.gitignore b/.gitignore index aa53365..9cdf251 100644 --- a/.gitignore +++ b/.gitignore @@ -37,3 +37,4 @@ downloads/ *.png !figure/*.R !figure-components/* +*.docx diff --git a/Workflow.Rmd b/Workflow.Rmd index ae7b874..17fd946 100644 --- a/Workflow.Rmd +++ b/Workflow.Rmd @@ -50,12 +50,11 @@ keywords: Single-cell RNA sequencing, expression variability, transcriptional noise, differential expression testing bibliography: Workflow.bib urlcolor: Orange -#output: -# BiocWorkflowTools::f1000_article: -# fig_width: 6 -# fig_height: 3.5 -output: - word_document +# output: +# BiocWorkflowTools::f1000_article: +# fig_width: 6 +# fig_height: 3.5 +output: word_document csl: american-medical-association.csl --- @@ -210,7 +209,7 @@ can be obtained from The `r Biocpkg("BASiCS")` Bioconductor package uses a Bayesian hierarchical model to simultaneously perform data normalisation, technical noise -quantification and downstream analyses [@Vallejos2015BASiCS;@Vallejos2016;@Eling2018] +quantification and downstream analyses [@Vallejos2015;@Vallejos2016;@Eling2018] within a cell population or populations under study. In this context, cell populations could correspond to groups set a priori by the experimental design (e.g. naive or stimulated CD4+ T cells in [@Martinez-jimenez2017]), @@ -1364,16 +1363,15 @@ ggplot(table_de) + ) ``` -While genes with significant changes in residual over-dispersion often have +Genes with significant changes in residual over-dispersion often have similar levels of mean expression, as seen in -Figure \@ref(fig:fig15-var-plots)A and C, -they may have a different proportion of zero counts in the two cell populations. -Figure \@ref(fig:fig15-var-plots)B and D show that many genes with higher residual -over-dispersion in somitic cells have a lower proportion of zeros in pre-somitic -mesoderm cells, -and vice versa. - -```{r fig15-var-plots, fig.width = 6, fig.height = 4, fig.cap="A, C: $\\log_2$ change in expression against log mean expression for genes with higher residual over-dispersion in somitic (A) cells and pre-somitic mesoderm (C) cells. B, D: Proportion of expressed cells for genes, with higher residual over-dispersion in somitic cells (B) and pre-somitic mesoderm (D) cells. Dashed red lines in panels A and C represent a log fold change of zero, meaning no change in average expression. Dashed red lines in panels B and D represent the line described by y=x, representing equal detection levels in both populations."} +Figure \@ref(fig:fig15-var-plots). +However, they may have a different proportion of zero counts in the two cell +populations, indicating a more bursting expression pattern, or similar proportion +of zero counts but more varying expression levels. We will now explore ways to +identify genes in each of these categories. + +```{r fig15-var-plots, fig.width = 6, fig.height = 4, fig.cap="$\\log_2$ change in expression against log mean expression for genes with higher residual over-dispersion in somitic (A) cells and pre-somitic mesoderm (B) cells. Dashed red lines represent a log fold change of zero, meaning no change in average expression."} ## mean expression vs log2FC for genes with higher residual over-dispersion in ## somitic cells g1 <- ggplot( diff --git a/Workflow.bib b/Workflow.bib index 1e03f5e..7776849 100644 --- a/Workflow.bib +++ b/Workflow.bib @@ -305,18 +305,6 @@ @article{Stegle2015 volume = {16}, year = {2015} } -@article{Vallejos2016a, -author = {Vallejos, Catalina A and Richardson, Sylvia and Marioni, John C}, -doi = {10.1101/035949}, -file = {:Users/nils/Documents/Mendeley Desktop/art{\%}3A10.1186{\%}2Fs13059-016-0930-3.pdf:pdf}, -journal = {Genome Biology}, -keywords = {cellular heterogeneity,differential expression,single-cell rna-seq}, -number = {70}, -title = {{Beyond comparisons of means: understanding changes in gene expression at the single-cell level}}, -url = {http://biorxiv.org/content/early/2016/01/05/035949.abstract}, -volume = {17}, -year = {2016} -} @article{Anders2010, author = {Anders, Simon and Huber, Wolfgang}, doi = {10.1186/gb-2010-11-10-r106}, @@ -647,19 +635,6 @@ @article{Ecker2017a url = {http://doi.wiley.com/10.1002/bies.201700148}, year = {2017} } -@article{Vallejos2015BASiCS, -author = {Vallejos, Catalina A. and Marioni, John C. and Richardson, Sylvia}, -doi = {10.1371/journal.pcbi.1004333}, -file = {:Users/nils/Documents/Mendeley Desktop/Vallejos, Marioni, Richardson - 2015 - BASiCS Bayesian Analysis of Single-Cell Sequencing Data.pdf:pdf}, -issn = {1553-7358}, -journal = {PLOS Computational Biology}, -number = {6}, -pages = {e1004333}, -title = {{BASiCS: Bayesian analysis of single-cell sequencing data}}, -url = {http://dx.plos.org/10.1371/journal.pcbi.1004333}, -volume = {11}, -year = {2015} -} @article{Vallejos2016, author = {Vallejos, Catalina A and Richardson, Sylvia and Marioni, John C}, doi = {10.1101/035949},