Extended Data Fig. 5: GSK3A regulates genome folding in a cohesin-dependent manner.
From: High-throughput Oligopaint screen identifies druggable 3D genome regulators
(a) Western blot of whole cell lysate after six hour auxin treatment in HCT-116-RAD21-mClover-AID cells and depletion of GSK3A after 72 h RNAi KD. Proteins are labelled with HRP-linked antibodies. All bands and lanes from same blot. Data shown represent one independent experiment. (b) Representative 3D DNA FISH images of chr22 domains following six-hour RAD21 auxin-inducible degradation, 72 h GSK3A KD or both. Dotted white line indicates nuclear edge. Scale bar for nucleus, 5 µm; scale bar for spots, 1 µm. Data shown represent one independent experiment. (c) Fold change in spatial overlap between chr22:D1-D2 after GSK3A KD, RAD21 degradation or both. ns P-value > 0.05, **** P-value < 0.0001, two-tailed Mann-Whitney U-test. [n = biologically independent alleles for Control (n = 669); GSK3A KD (n = 712); +Auxin (n = 680); GSK3A KD + Auxin (n = 560)]. (d) Representative 3D DNA FISH images of chr22 domains after auxin-inducible degradation of RAD21, 24 h treatment with BRD0705 (GSK3Ai) or both. Dotted white line indicates nuclear edge. Scale bar nucleus, 5 µm; Scale bar spots, 1 µm. Data shown represent one independent experiment. (e) Fold change in mean spatial overlap at chr22:D1-D2. ns P-value > 0.05, * P-value < 0.05, **** P-value < 0.0001, two-tailed Mann-Whitney U-test. [n = biologically independent alleles for control (n = 350), BRD0705 (n = 302), +Auxin (n = 525), BRD0705+Auxin (n = 280)].
