Extended Data Fig. 4: Characterization of the WapldTagCdca5mAID and WapldTagSmc2mAIDCdca5mAID cell lines.
From: Extensive mutual influences of SMC complexes shape 3D genome folding
a, Schematic showing the edited Cdca5 (encoding Sororin), Wapl and Smc2 loci and the respective genotyping strategy. b, Genotyping results of the WapldTagCdca5mAID clones. One experiment was performed. c, Fluorescence images showing loss of WAPL and Sororin fluorescence signals upon 4 h of dTag13 and 5-Ph-IAA treatment in the WapldTagCdca5mAID cells. Two independent clones were tested. Scale Bar: 10 μm. d, Schematic showing the strategy to determine G2/M arrest length. e, Metaphase spread analysis showing a more extensive dissociation of sister-chromatids in the WapldTagCdca5mAID cells after 16 h compared to 6 h of RO-3306 mediated G2 arrest. Scale Bar: 10 μm. Four independent experiments were performed. f, Quantification of (d & e), showing progressive dissociation of sister-chromatids as G2-arrest duration were extended. Longer G2-arrest allowed more complete removal of cohesive-cohesin by WAPL. Data are presented as mean values +/- SEM (n = 4 independent experiments for all time points). P values were calculated by two-sided student’s t-test. g, Genotyping results of WapldTagSmc2mAIDCdca5mAID clones. One experiment was performed. h, Fluorescence images showing loss of WAPL, Sororin and SMC2 fluorescence signals upon 4 h of dTag13 and 5-Ph-IAA treatment in the WapldTagSmc2mAIDCdca5mAID cells. Three independent clones were tested. Scale Bar: 10 μm.
