Dynamics of LC Ig-fold laminopathy mutants and validation of BAF-KD. (A) Dynamics of mEmerald-LC Ig-fold point mutants in Lmna-KO MEF. The right four columns are magnified views of orange boxes, and the rupture sites are indicated with yellow arrowheads. Bars: 5 μm (the first column) and 2 μm (the second to fifth column). (B and C) Validation of BAF-KD with immunofluorescence (B) and immunoblotting (C). (B) Representative immunofluorescence images of single confocal sections in WT MEFs expressing scrambled control (shScr), shBAF#1 or shBAF#2 with sfGFP-DARPin-LA6 and sfCherry stained with anti-BANF1/BAF (EPR7668), followed by Cy5-labeled anti-rabbit IgG, and Hoechst 33342 for DNA. Bar: 20 μm. (C) Whole cell lysates from MEFs expressing the indicated shRNAs were probed with anti- BANF1/BAF (EPR7668) and anti-GAPDH (as loading control). Positions of the size standards are shown on the right. (D) Representative images of single confocal sections of sfGFP-DARPin-LA6 in a NE protrusion in MEFs fixed within 10 min after microirradiation. DNA was stained with Hoechst 33342. The right image of each column is magnified view of orange box. The edges of protruded DNA regions are indicated with yellow dotted line (top right of each column). Bars: 5 μm (the left of each column) and 2 μm (the right of each column). Source data are available for this figure: SourceData FS4.