Extended Data Fig. 9: Characterization of the G1E-ER4:mCherry-mAID-HP1α;GFP-FKBPF36V-HP1β cell line.
From: Genome folding principles uncovered in condensin-depleted mitotic chromosomes
a, Schematic illustration showing the homozygous insertion of mCherry-mAID tag and GFP-FKBPF36V tag to the N terminus of endogenous Cbx5 and Cbx1 locus, respectively. b, Western blot showing the degradation of HP1α and HP1β upon 5-Ph-IAA or/and dTag13 treatment. One experiment was performed. Loading control was processed on a different gel. c, Schematic illustration showing the strategy of nocodazole-based arrest/release in conjunction with 5-Ph-IAA or/and dTag13 treatment. Early and late-G1 phase cells with four distinct HP1 protein configurations were collected. d, Flow cytometry plot showing the sorting strategy of early-G1 and late-G1 phase cells with distinct HP1 protein configurations. Two biological replicates were performed. e, Representative confocal images showing the successful depletion of HP1α or HP1β or both in the sorted G1 phase cells. Scale bar: 10 μm. Two biological replicates were performed. f, Flow cytometry plot showing the mitotic progression of cells under distinct configurations of HP1 proteins after nocodazole release. Two biological replicates were performed.
