Supplementary Figure 5: TRR facilitates decatenation of UFBs by Top2a.
From: PICH and TOP3A cooperate to induce positive DNA supercoiling
(a) Western blot analysis of Top3a, PICH and Top2a in HCT116-Top3aAID (Top3dg) and in the control cell line (430) in metaphase arrested cells incubated for 3 h in the absence (-) or in the presence (+) of Auxin. For each panel, representative images out of at least 3 independent experiments are presented. Uncropped blots images are shown in Supplementary Data Set 1. (b) Quantification of the average number of UFBs in the control cell line (430) exposed (red bars) or not (gray bars) to Auxin. Cells were treated with solvent (-) or either 50 nM or 100 nM ICRF193, as indicated below. Error bars denote the standard deviation of 3 independent replicates (n = 80cells/replicate) (c) Additional representative immunofluorescence images of HCT116-Top3aAID cells following Top3a depletion (+Auxin) in cells progressing through anaphase in the absence of ICRF193. UFBs are marked by PICH (blue) and centromeres are marked by CREST (red). The zoomed image shows CREST-positive staining at the tips of UFBs. Scale bar is 10 µm. Representative images of at least 20 images collected for 3 independent replicates are presented.
