Fig. 7: Ph with an intact SAM increases H2A-ub in vivo. | Nature Communications

Fig. 7: Ph with an intact SAM increases H2A-ub in vivo.

From: Phase separation by the polyhomeotic sterile alpha motif compartmentalizes Polycomb Group proteins and enhances their activity

Fig. 7: Ph with an intact SAM increases H2A-ub in vivo.The alternative text for this image may have been generated using AI.

a Representative Western blot of Ph levels in induced cell lines. Note that Ph-ML is the strong double mutant (L1547R/H1552R), which was previously shown to disrupt Ph clustering in cells10. b Representative Western blot of histone H2A-Ub levels in induced cell lines. Blots were reprobed with anti-H3 to normalize loading. In total, 125 and 250 ng of acid-extracted histones were loaded for each sample. c, d Quantification of Ph (c) and H2A-Ub (d) for three experiments. p Values are for one-way ANOVA comparing Ph-WT and Ph-ML cells to control (S2) for the 250 ng point. e–g Representative images of cells overexpressing Venus-Ph (e), Venus-Ph-ML (f), or Venus-PhΔSAM (g) by transient transfection under control of the heat-shock promoter. Scale bar shown in (g) applies to (e–g). h Graph of the number of foci per cell for two independent experiments. Note that only cells with >zero foci were included. p Values are for Kruskal–Wallis tests with Dunn’s multiple-comparison correction. Comparisons were made among cells expressing different versions of Ph for both experiments. n Values (# cells analyzed in experiments 1 and 2) in Venus-Ph 118, 144; Venus-Ph-ML 128, 63; Venus-PhΔSAM 72, 30. See also Supplementary Fig. 18.

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