Fig. 2: s-ISR is positively regulated by eIF4E.
From: Plasticity of the mammalian integrated stress response
a, Absorbance profiles (254 nm) of 10–50% sucrose gradients from indicated cell lines. b–d, Scatter plots comparing fold change (FC) in total and polysome-associated mRNA quantified using RNA sequencing in shEif2b5 versus shCon (b), shEif2b5 + shEif4e versus shEif2b5 (c) and Tg 16 h versus control6 (d) MEFs. Differentially regulated genes are colour-coded (legend under d). All genes are shown in Supplementary Table 1. e–p, Comparisons of how genes controlled at the level of mRNA abundance (e–j) or translation (k–p) are regulated in shEif2b5 versus shCon (e–g,k–m) and shEif2b5 + shEif4e versus shEif2b5 (h–j,n–p) MEFs. For each gene set and comparison, a scatter plot (e,h,k,n) and empirical cumulative distribution plots for log2[FC] in total (f,i,l,o) and polysome-associated (g,j,m,p) mRNA are shown. Gene sets are colour-coded as indicated in d, and background (non-affected) genes are shown in grey. Shifts in the distribution of fold changes for gene sets relative to the background were assessed for total (f,i,l,o) and polysome-associated (g,j,m,p) mRNA using the Mann–Whitney U-test and indicated P < 0.01 for all comparisons. q, Gene Ontology (GO) enrichment analysis for regulated genes (through mRNA abundance or translation) from b. The number of genes (No.) identified in selected GO pathways is indicated, and all GO pathways are shown in Supplementary Table 2. r, Proliferation of MEFs expressing the indicated shRNAs was determined by counting live cells (trypan blue exclusion). P value was determined by the two-tailed Student’s t-test. Data are presented as mean ± s.e.m. Inset shows representative western blot analysis of levels of the indicated proteins. (n = 3 independent experiments).
