Fig. 3: Reduced snRNP production upon transcriptional inhibition.
From: Interaction of 7SK with the Smn complex modulates snRNP production
a Western blot analysis of Smn and Larp7 protein levels in NSC-34 cells treated with DMSO or ActD for the indicated durations. b Quantification of Larp7 protein levels in a. Data are mean ± s.d.; ****P ≤ 0.0001; n.s., not significant; two-way ANOVA with Sidak’s multiple-comparisons test (n = 3). c Co-immunoprecipitation of Larp7 by anti-Smn from NSC-34 cells treated with DMSO or ActD for the indicated durations. d Quantification of Larp7 co-purification by anti-Smn in c. Data are mean ± s.d.; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; n.s., not significant; two-way ANOVA with Sidak’s multiple-comparisons test (n = 3). e Co-immunoprecipitation of Mepce by anti-Smn from NSC-34 cells treated with DMSO or ActD for 6 h. f Co-immunoprecipitation of Mepce and Smn by anti-SmB/B’ from NSC-34 cells treated with DMSO or ActD for 6 h. Note that the immunosignal at 25 kDa for mouse-IgG immunoprecipitation is non-specific from the antibody light chain. g qPCR analysis of total snRNA levels in NSC-34 cells treated with DMSO or ActD for the indicated durations. Data are mean ± s.d.; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; two-way ANOVA with Sidak’s multiple-comparisons test (n = 3). h qPCR analysis of snRNAs co-precipitated by anti-SmB/B’ from NSC-34 cells treated with DMSO or ActD for the indicated durations. Data are mean ± s.d.; *P ≤ 0.05; ****P ≤ 0.0001; two-way ANOVA with Sidak’s multiple-comparisons test (n = 3). i Agarose gel electrophoresis of biotinylated U2 snRNA. j In vitro snRNP assembly assay. Biotinylated U2 snRNA was incubated with cytosolic extracts from NSC-34 cells exposed to DMSO or ActD for 6 h. Reactions were assembled with or without exogenous ATP. Biotinylated U2 snRNPs were pulled down with streptavidin beads and SmB/B’ was analyzed by Western blot. k Quantification of in vitro U2 snRNP assembly assay in j. Data are mean ± s.d.; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; two-way ANOVA with Tukey’s multiple-comparisons test (n = 3). Source data are provided as a Source Data file.
