Figure 4.

Actin Accumulation Causes Nuclear Deformation

(A) The nuclear envelope of keratinocytes (left panel) and siNT- or siCFL1-treated ADF-null SCCs (right panel) was stained for Lamin A/C. The scale bar represents 20 μm.

(B) 3D model construction composed of nuclei (blue) and F-actin (red) of cells treated as in (A). Model constructed using IMARIS software (Bitplane) after 0.4-μm optical slices were obtained using confocal microscopy. The siNT scale bar represents 7 μm and siCFL1 scale bars represent 10 μm, 20 μm, and 10 μm (left to right images).

(C) Images of ADF-null SCCs treated with siNT or siCFL1 were obtained by structured illumination microscopy (SIM). Cells were stained for Lamin A/C. Images were taken as part of a z stack, available as Figure S3A. The scale bar represents 10 μm.

(D) IF staining for β-actin (left panels) and Lamin A/C (right panels) performed on skin sections. Zoomed images of boxed region are shown below panels. The scale bars represent 20 μm.

(E) Nuclear size quantification based on Lamin A/C staining of 88 control and 183 double knockout nuclei (n = 2) performed in ImageJ. All individual measurements are presented, and bars represent mean ± SD. ^∗∗∗Mann-Whitney p value < 0.0001.

Nuclei and F-actin were counterstained with DAPI and phalloidin, respectively. See also Figure S3.