Supplementary Figure 1: Chromatin deposition of macroH2A variants is replication independent (related to Fig. 1).
From: Transcription-associated histone pruning demarcates macroH2A chromatin domains
(a) Western blot analysis of whole cell lysate showing the expression of SNAP-tagged histones in NIH 3T3 cells. Arrowheads indicate SNAP-tagged proteins and asterisks indicate endogenous proteins. Amido black staining of histones used as loading control. (b) Profiles of cell cycle distribution for SNAP-H3.1 or macroH2A2-SNAP in NIH 3T3 cells based on DNA content. Numbers represent percentage of nuclei in the indicated cell cycle phase. (c) FACS analysis showing quantification of median SNAPc in NIH 3T3 cells stably expressing SNAP-H3.1 or macroH2A2-SNAP with indicated labeling strategies as well as parental cells. (d) Same as in (a) in WT iDFs. (e) qPCR analysis of WT iDFs in distinct cell cycle stages sorted based on DNA content, showing the mRNA expression level of endogenous macroH2A1.1, macroH2A1.2 and macroH2A2. E2f1 (G1), Pcna (G1), H4 (S) and Cenpa (G2) are used as controls. Rpl7 is used as internal reference for normalization. Error bars represent s.d. from n = 2 independent experiments. P values are calculated from two-tailed t test comparing cell cycle enriched fractions with the asynchronous population (*P < 0.05, ** P < 0.01, *** P < 0.001).
