Figure 5

CTDNEP1 loss increases rupture frequency in the absence of NPC or confinement defects. (a) CTDNEP1 depletion by single siRNAs increases nucleus rupture frequency in U2OS-shLMNB1 RFP-NLS and HeLa GFP-Nuc cells compared to control, analyzed by live-cell imaging. (b) CTDNEP1 depletion does not significantly increase rupture duration, analyzed by live-cell imaging in U2OS-shLMNB1 RFP-NLS. (c) Micronucleus rupture frequency analyzed in fixed asynchronous cells induced to form micronuclei via spindle assembly checkpoint inhibition. (d) Representative DAPI orthosections of cells treated with indicated siRNAs showing similar nuclear height profiles. (e) Quantification of nuclear height after single siRNA transfections shows no significant change after CTDNEP1 depletion. (f) Representative images of actin organization (phalloidin) in siRNA treated cells. Max. intensity projections of cell top half. (g) Representative images of NUP133 foci. Max. intensity projection of nucleus top half in siRNA-treated cells. (h) NUP133 quantification showing decreased foci density following CTDNEP1 depletion. Cells: U2OS GFP-Nuc, unless indicated. Stats: Tables S14–S18. For all graphs, N = 3, *p < 0.05, ns = p > 0.05. For (a) and (c): Barnards test. For (b,e,h): K-S test.