Extended Data Fig. 7: GSK3A genetically interacts with WAPL to regulate cohesin levels.
From: High-throughput Oligopaint screen identifies druggable 3D genome regulators
a) Plasmid maps for inserting WAPL-mAID2-mClover3-Hygromycin and WAPL-mAID2-mClover3-Neomycin into parental HCT-116-OsTir1(F74G) cell line to create HCT-116-WAPL-AID2 cell line. DNA electrophoresis below confirms insert of both cassettes. b) Western blot of WAPL from HCT116-WAPL-AID lysate +/− WAPL KD and +/− auxin-inducible degradation of WAPL. LE = long exposure, SE = short exposure. Data shown represent one independent experiment. c) Representative images of RAD21 immunofluorescence after auxin-induced degradation of WAPL and/or siRNA treatment. Median RAD21 signal granularity noted in red. Quantification in Extended Data Fig. 5. Scale bar nucleus, 5 µm. d) Median RAD21 signal granularity for +/− siRNA treatment and +/− degradation of WAPL. Each bar is median. [Replicate wells for -Auxin, Control (n = 13), GSK3A (n = 14), WAPL (n = 9), GSK3A + WAPL (n = 14). For +Auxin, Control (n = 12), GSK3A (n = 14), WAPL (n = 12), GSK3A +WAPL (n = 14)] **** P-value < 0.0001, two-tailed t-test.
