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. 2020 Jan 30;11:612. doi: 10.1038/s41467-020-14511-w

Fig. 4. H3R2me2a acts as a histone mark for CPC recruitment on chromosome arms.

Fig. 4

a–d The cells were treated with siPRMT6, MS023 as a PRMT6 inhibitor, or CHR-6494 as a Haspin kinase inhibitor. The CPC/CREST intensity ratio was determined by immunofluorescence microscopy of nocodazole-arrested HeLa chromosome spreads at 100 centromeres and 100 chromosome arms per condition from three independent experiments. e The cytosol and chromosome fractions were isolated by centrifugation from mitotic HeLa cells treated with Haspin or a PRMT6 inhibitor and analyzed by Western blot probing for the indicated proteins. Histone H3 and tubulin served as loading controls for chromosome and cytosol fractions, respectively. f, g The cells were treated with siPRMT6, PRMT6 inhibitor, or Haspin kinase inhibitor. The average intensity of H3S10ph in individual chromosomes was determined by immunofluorescence microscopy of nocodazole-arrested HeLa chromosome spreads as in (b) and was plotted (g, n = 100 chromosomes from three independent experiments). Scale bars, 5 μm. Error bars, SEMs. P values were calculated by two-tailed Student’s t-tests (b and c; *p < 0.01) or two-way ANOVA (g; *p < 0.01). Source data are provided as a Source Data file.