Difference of lamin isoforms in the accumulation kinetics at the rupture sites in MEFs, C2C12, BJ-5ta and MCF10A cells. (A–D) A 2-μm diameter spot at the NE in WT MEFs (A), C2C12 (B), BJ-5ta (C), and MCF10A (D) were laser-microirradiated to induce the rupture, fixed within 10 min (left panel of each) or 60–70 min later (right panel of each), and then stained with a combination of anti-mouse and anti-rabbit antibodies, followed with Alexa Fluor 488-labeled anti-rabbit IgG and Cy5-labeled anti-mouse IgG, and Hoechst 33342 for DNA. Magnified views of the indicated areas with orange boxes are shown (the second to fifth columns). The ruptured sites are indicated with yellow arrowheads (the second columns). Representative images of single confocal sections. Color-merged images (the first and second columns) in (A) MEFs show anti-LC (ab125679, green)/anti-LA (C-3, magenta), (B) C2C12 cells show anti-LC (321, green)/anti-LA (C-3, magenta), and anti-LB2 (EPR9701(B), green)/anti-LB1 (B-10, magenta), (C) BJ-5ta cells show anti-LC (321, green)/anti-LA (C-3, magenta), and anti-LB2 (EPR9701(B), green)/anti-LB1 (8D1, magenta) and (D) MCF10A cells show anti-LC (321, green)/anti-LA (C-3, magenta), and anti-LB2 (EPR9701(B), green)/anti-LB1 (8D1, magenta). Bars: 5 μm (the first column) and 2 μm (the second to fifth column). (E) Ratios of cells with (green) and without (gray) enrichments of the indicated antibodies at the rupture sites. The numbers of analyzed cells, fixed within 10 min (left panel) and 60 min (right panel) after laser microirradiation are indicated in the bar charts.