Ab16048

The nuclear morphologies of the SK-N-SH cells were visualized by immunostaining with lamin A and lamin B1. The lamin A and vimentin levels in the cells were characterized by immunostaining of lamin A and vimentin. Cells were cultured on nanopillars and incubated overnight before fixation with prewarmed 4% paraformaldehyde (PFA) in PBS (Boster biological technology AR1068) for 15 minutes at room temperature. After washing with PBS three times, the cells were permeabilized in 0.5% Triton X-100 (Sigma) in PBS for 15 minutes, followed by blocking with 5% bovine serum albumin (BSA) (Sigma) in PBS for 1 hour. Subsequently, the samples were incubated with primary antibodies (anti-lamin A, Abcam, ab26300; anti-lamin B1, Abcam, ab16048; anti-vimentin, Sigma, V5255; anti-SNAIL + SLUG, Abcam, ab224731) at 1 : 400 dilution at room temperature for 1 hour or 4 °C overnight. The samples were again washed with PBS for three times and then incubated with secondary antibodies (chicken anti-rabbit IgG Alexa 488, Invitrogen, A21441; anti-mouse IgG Alexa 555, Cell Signaling Technology, 4409s) at 1 : 600 dilution at room temperature for 1 hour. After washing with PBS three times, the samples were stained with DAPI (Sigma), phalloidin (Cytoskeleton, Inc.) or cellmask (Invitrogen) in different experiments.