H3K36me2 is enriched in PCH in some mouse cells. (A–C) Localization of H3K36me2 by immunofluorescence. Pmi28 cells (A) and iMEFs (B) were fixed and stained with anti-H3K36me2 (green), anti-H3K9me3 (red), and Hoechst 33342 (blue). (A and B) Single optical sections of confocal microscope images are shown. Insets show magnified views of indicated areas. Line plots indicate intensity profiles of H3K36me2, H3K9me3, and DNA along the yellow line in the merged images. Contrast was adjusted individually to compare the localization patterns. (C) Quantification of the intensity ratio of H3K36me2 in PCH to that in the nucleoplasm in various mouse cell lines. Two box plots display data from two biological replicates with dots representing individual cells (round for the first experiment and triangular for the second). Center lines represent medians; box limits indicate the 25th and 75th percentiles; whiskers extend 1.5 times the interquartile range from these percentiles. The numbers of total cells analyzed (n) and P value comparing ESCs (H3K36me2 in PCH) to MC12 cells (H3K36me2 in euchromatin), calculated using the Mann–Whitney U test, are indicated. (D) Genome-wide correlation heatmap of H3K36me2 and H3K9me3 in mouse ESCs, B cells, and pancreatic cancer cells. Spearman’s correlation coefficients are indicated inside cells. Scale bars, 10 μm.