FIGURE 4:

Lamin A/C meshwork defects are reversed by actin depolymerization. (A) Overview of experimental strategy. mNG-AID-LB1 cells were treated with DMSO or auxin for 16 h followed by an hour of DMSO or Lat A treatment. mNG-AID-LB1 cells were then fixed and immunostained. (B) xz view to show nuclear height of mNG-AID-LB1 cells immunostained for lamin A/C after 16 h of DMSO or auxin treatment followed by 1 h of additional DMSO or Lat A. (C–F) Immunofluorescence of mNeonGreen, lamin A/C and actin after mNG-AID-LB1 cells were treated with (C) DMSO or (E) auxin for 16 h. mNG-AID-LB1 cells were then treated with (D) Lat A for an hour or (F) 16 h of auxin followed by 1 h of Lat A treatment; scale bar, 10 µm. (G) Quantification of the percentage of nuclei with lamin A/C defects (detected by blinded categorical scoring) after mNG-AID-LB1 cells were treated with DMSO or auxin for 16 h followed by 1 h of additional DMSO or Lat A treatment. (H) Normalized histograms of the percentage of lamin A/C-free nuclear surface area in mNG-AID-LB1 cells after 16 h auxin treatment (blue) or 16 h auxin treatment + 1 h Lat A treatment (gray). Curves indicate mean and error bars indicate SD of three independent experiments. (I) Summary plot of the percentage of nuclei with contiguous lamin A/C (<1% lamin A/C-free nuclear surface, gray), with gaps in the lamin A/C meshwork covering >1% of the nuclear surface (orange), or with gaps covering >10% of the nuclear surface (red) in mNG-AID-LB1 cells in the indicated conditions. n > 400 cells analyzed per condition in three independent replicate experiments. ****, p < 0.0001 by the Fisher's exact test.