Extended Data Fig. 8: Increase in Raf1 abundance promotes caffeine resistance through heterochromatin formation.
From: Stress controls heterochromatin inheritance via histone H3 ubiquitylation
a, b, Real-time qPCR analysis of indicated genes in cells treated with 16 mM caffeine for 2 weeks in WT or raf1R576H cells. Data are presented as mean ± SD; n = 3 independent experiments, P value by two-stage step-up Benjamini, Krieger, and Yekutieli unpaired t test for (a) and one-way ANOVA followed by Tukey’s multiple comparisons test for (b). c, Western blot analysis of Raf1 in raf1R576H cells treated with 16 mM caffeine for 2 weeks. Cdc2 and Ponceau S staining included as loading controls. d, Serial dilutions of the indicated strains were plated on nonselective (N/S) or 16 mM caffeine-containing medium. e, Quantification of caffeine-resistant colonies in raf1-oe cells (Padh1-raf1) with or without clr4 (data are presented as mean ± SD; n = 3 independent experiments, P value by two-tailed unpaired t test). f, ChIP-seq analyses of H3K9me3 enrichment in cells treated with 16 mM caffeine for 2 weeks, with or without raf1-oe. Facultative heterochromatin islands with increased H3K9me3 levels in raf1-oe cells are indicated. g, Representative plots of H3K9me3 distribution at facultative islands responsive to caffeine. Open reading frame (ORF) positions are indicated. Data are representative of two independent experiments. Source data are provided.
