FIGURE 7:

LAP2β and LB1 codepletion induces nuclear rupture in an actin-dependent manner. (A) Overview of experimental strategy. mNG-AID-LB1 cells were subject to RNAi for 48 h followed by 16 h of DMSO or auxin treatment and live imaging at ×40 magnifications with images of 3xNLS-TdTomato and H2B-IRFP acquired every 3 min for 4 h. (B–E) Still images from time-lapse imaging of mNG-AID-LB1 cells expressing 3xNLS-tdTomato and treated with control RNAi (B), control RNAi + auxin(C), LAP2β RNAi (D), or LAP2β RNAi + auxin (E). See also Supplemental Movies S4 and S5. (F) Overview of experimental strategy for panels (G–J). mNG-AID-LB1 cells were subject to RNAi for 48 h followed by 16 h of DMSO or auxin treatment and 1 h of either Lat A drug treatment (G–I) or colchicine drug treatment (J) followed by live imaging at ×40 magnifications with images of 3xNLS-TdTomato and H2B-mIRFP acquired every 3 min for 4 h. (G and H) Still images from time-lapse imaging of mNG-AID-LB1 cells expressing 3xNLS-tdTomato and treated with LAP2β RNAi and Lat A (G) or LAP2β RNAi + auxin + Lat A (H); scale bar, 20 µm. See also Supplemental Movies S7 and S8. (I) Quantification of NE rupture in AID-LB1 cells after indicated RNAi, 16 h of DMSO or auxin followed by additional DMSO or Lat A. Each dot on graph represents one replicate; n > 200 cells analyzed per condition. (J) Quantification of NE rupture in AID-LB1 cells after indicated RNAi, 16 h of DMSO or auxin followed by additional DMSO or colchicine. See also Supplemental Movie S9. Each dot on graph represents one replicate; n > 420 cells analyzed per condition. ns indicates p > 0.05; **, p < 0.01; and ****, p_adj < 0.0001 by the unpaired t test.