Extended Data Fig. 3: Splicing factor mutation mimics NMD loss in causing raf1 upregulation.
From: Stress controls heterochromatin inheritance via histone H3 ubiquitylation
a, RNA-seq splice junctions reveal cryptic introns in raf1. Numbers indicate the percentage of spliced reads. b, Mutation in the Sap49 splicing factor or the NMD component Upf2 suppresses the heterochromatic silencing defect in raf1R576H cells. Serial dilutions assessed mat2P::ura4+ expression, and iodine staining assessed haploid meiosis (top). Domain organizations of Sap49 and Upf2 are shown (bottom). c, d, e, ChIP-seq profiles of H3K9me3 across the silent mat region (c), the subtelomeric region 1R (d), and centromere 2 (e) in the indicated strains. f, Northern blot of raf1 transcripts in the indicated strains; ribosomal RNA served as the loading control. g, h, Western blot analysis of Raf1 in the indicated strains. Cdc2 and Ponceau S served as loading controls. i, Expressing Raf1 in raf1-oe (Pnmt1-raf1) cells at levels comparable to those in upf1Δ cells is sufficient to suppress the heterochromatic silencing defect in raf1R576H cells. Serial dilutions on the indicated media were used to assess mat2P::ura4+ expression. Data are representative of two independent experiments. Source data are provided.
