Fig. 5: ERAS is conserved in mammalian cells.
From: ER-associated RNA silencing promotes ER quality control
a,b, RNA sequencing results of tunicamycin/DMSO-treated (a) and thapsigargin/DMSO-treated (b) Ago2+/+ and Ago2−/− MEF cells. Ago2-specific targets were selected by filtering (P ≤ 0.05) transcripts that were downregulated in Ago2+/+ and upregulated in Ago2−/− cells (cyan). c, Venn diagram showing the number of transcripts regulated by AGO2 upon ER stress induction. Only transcripts that were identified in both ER stress conditions (a and b) were further investigated (cyan). d, Cellular component GO term enrichment of the genes identified in a and b. e, Expression levels of endogenous Nov, Mertk and C1rl mRNA in tunicamycin-treated MEFs measured by qRT–PCR. Data relative to 18S rRNA. f, UV CLIP of tunicamycin-treated MEF cells using AGO2 as bait and subsequent qRT–PCR of target mRNAs in the input (IN), supernatant (SN) and immunoprecipitation (IP) fractions. Data normalized to input fraction. Top shows western blot against AGO2 for the performed CLIP experiment. g, Dose-dependent cell survival after tunicamycin treatment calculated from the colony formation assay shown in Extended Data Fig. 5f. Bar graph showing the IC50 values for tunicamycin between Ago2+/+ and Ago2−/− MEF cells. h–k, α-Amanitin chase assays for indicated endogenous mRNAs identified in a and b from Ago2+/+ and Ago2−/− MEF cells pre-treated with tunicamycin. mRNA measured by qRT–PCR. Data relative to 18S and 28S rRNA. Data in e were analysed by unpaired two-tailed t-tests. Data in f were analysed by a two-way ANOVA with Dunnett’s multiple comparison test. The bar graph in g was analysed by paired two-tailed t-test. In g–k, data were analysed by two-way ANOVA with Sidak´s multiple comparison test. In e-k values are depicted as mean ± s.e.m. generated from n = 3 independent experiments, *P < 0.05; **P < 0.01; ***P < 0.001; NS, P > 0.05. In d, raw P values were corrected for FDR. *FDR <0.05; ** FDR <0.01; *** FDR <0.001; NS, FDR >0.05. Source numerical data and unprocessed blots are available in source data.
