Fig. 6.

Depletion of SMYD5 leads to decreased H4K20me3, H3K9me3/2, and HP1α binding. a Change in the global distribution of H4K20me3 in shSmyd5 ES cells. b Average profile and boxplot of H4K20me3 ChIP-Seq tag density in shSmyd5 ES cells. c Average profiles of H4K20me3 at SMYD5-enriched regions in shLuc and shSmyd5 ES cells. d, e Changes in global distributions of d H3K9me3 (e) and H3K9me2 in shSmyd5 ES cells relative to shLuc ES cells. f–h Average profiles and boxplot of f H3K9me3, g H3K9me2, and h HP1α densities in shLuc and shSmyd5 ES cells. i Boxplot depicting density of H4K20me3, H3K9me3, and H3K9me2 at H4K20me3 islands. j Empirical cumulative distribution (ECDF) for the fold-change in density of HP1α in shSmyd5 ES cells. The red line shifted to the left of the input (gray) shows a systematic decrease in enrichment in shSmyd5 ES cells. Boxplot below shows HP1α density (log2 fold-change vs. input) in shLuc and shSmyd5 ES cells. k H4K20me3 density at regions at regions with decreased or unaltered H3K9me3 levels. l SMYD5 associates with HP1α and G9a. FLBIO-SMYD5 (biotinylated SMYD5 + BirA) or BirA (control) ES cells were used to immunoprecipitate SMYD5 protein with avidin-agarose beads. Immunoprecipitates were analyzed by immunoblotting with anti-HP1α, anti-G9a, and anti-SMYD5 antibodies. m Changes in the global distribution of G9a in shSmyd5 ES cells relative to shLuc ES cells. n Altered profiles of H4K20me3 at SMYD5-enriched regions (H2-Q1, H2-Q7) in shLuc and shSmyd5 ES cells