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. Author manuscript; available in PMC: 2023 Mar 24.
Published in final edited form as: Nat Cell Biol. 2023 Jan 5;25(1):42–55. doi: 10.1038/s41556-022-01051-2

Extended Data Fig. 1: Characterization of WT and mutant CiA Oct4 dual reporter cells.

Extended Data Fig. 1:

a) Western blot confirms expression of TetR-FLAG and TetR-FLAG-HP1 proteins in WT and Dnmt1 knockout (KO) CiA Oct4 dual reporter ESCs. LMNB1 is used as protein loading control. b) FACS gating strategy used to analyse the GFP fluorescence in cells. c) Flow cytometry histograms show GFP expression in CiA Oct4 dual reporter cells after TetR-FLAG recruitment and after TetR-FLAG release following Dox addition for four days. d) ChIP-qPCR shows relative enrichment of HP1γ upstream of the TetO site before tethering, in the presence of TetR-FLAG-HP1 and after Dox-dependent release of TetR-FLAG-HP1 for four days. n = 2 independent biological replicates. e) Bar plot shows fraction of cytosine methylation (5mC) in WT and Dnmt1 KO CiA Oct4 dual reporter ESCs measured by LC-MS. n = 3 independent biological replicates. Data are presented as mean values +/− SD. f) Western blot shows expression of TetR-FLAG-HP1 and ZFP462 in WT and two independent Zfp462 KO CiA Oct4 dual reporter ESC lines. LMNB1 is used as loading control.