The effect of re-expression of A-type lamins. (A) Validation of A-type lamins re-expression in Lmna-KO MEFs. Whole cell lysates from MEFs expressing the indicated constructs were probed with anti-LA/C and anti-GAPDH (as loading control). PB533-mEmerald-C1-Neo were used as control (empty). Positions of the size standards are shown on the right. The degradation fragment is indicated with #. The contribution of the fragment to rescue effects is likely to be marginal because the expression level is significantly low compared to mEmerald-LA (top band). (B–E) Representative images of single confocal sections of the indicated MEFs fixed within 10 min after laser microirradiation and stained with anti-BANF1 (3F10-4G12; B) or anti-cGAS (D), Cy5-labeled anti-mouse IgG or rabbit IgG, and Hoechst 33342 for DNA. The max intensities of BAF (C) and cGAS (E) signals at the rupture sites in the indicated MEFs. The plotted points are from two independent experiments (n = 30, 29, 32, and 32 for Empty, LA, LC, and LA + LC, respectively; significance was determined by a Kruskal–Wallis test, P < 0.05; far outliers were excluded if P < 0.001 by the Smirnov-Grubbs test; horizontal dotted lines show the mean values, *, P < 0.05 from empty control, †, P < 0.05 from LC by a Steel–Dwass multiple comparison after far outliers were excluded). (B and D) Bars: 20 μm. Yellow arrowheads indicate laser microirradiation-induced NE rupture sites. Blue arrowheads indicate spontaneously produced NE protrusions. Source data are available for this figure: SourceData F9.