Fig. 2: Post-transcriptional regulation of ERAD substrate and viral RNA level.
From: ER-associated RNA silencing promotes ER quality control
a,b, CHX chase assay to assess CPL-1* protein stability: western blot of lysates from cpl-1* expressing worms with indicated mutations detecting CPL-1* (YFP) and tubulin (TUB) over 9 h of CHX treatment (a); BioSorter quantification of YFP fluorescence in vehicle (EtOH)- and CHX-treated cpl-1*-expressing worms (b). CHX/EtOH ratios were normalized to the 1 h timepoint of the respective genotype. c, α-Amanitin chase assay to monitor cpl-1 mRNA level in rde‑1(ne219) and drh‑1(ok3495) mutant worms expressing cpl-1*. Northern blot of purified RNA from animals expressing cpl-1*, detecting cpl-1 mRNA and 5.8S rRNA. 28S and 18S rRNA served as agarose gel loading control. d, Quantification of cpl-1 mRNA level corresponding to the data shown in c, relative to 5.8 S rRNA. e, α-Amanitin chase assay to monitor cpl-1 mRNA level in rde‑1(ne219) and drh‑1(ok3495) mutant worms expressing cpl-1WT. Northern blot of purified RNA from animals expressing cpl-1WT, detecting cpl-1 mRNA and 5.8 S rRNA. 28S and 18S rRNA served as agarose gel loading control. f, Quantification of cpl-1 mRNA level corresponding to the data shown in e, relative to 5.8S rRNA. g, Viral RNA1 level measured by qRT–PCR in worms infected with the Orsay virus. h, Viral RNA1 level measured by qRT–PCR in worms pre-treated with vehicle (DMSO) or tunicamycin before viral infection. Data are relative to 18S rRNA. i, Viral RNA1 level measured by qRT–PCR in worms, defective for the UPRER pathway. In b, d and f–i, Values are depicted as mean ± standard error of the mean (s.e.m.) generated from n = 3 independent experiments, *P < 0.05; **P < 0.01; ***P < 0.001; NS, P > 0.05. Data were analysed by two-way ANOVA with Sidak’s multiple comparison test. Source numerical data and unprocessed blots are available in source data.
