For lamin staining, cells were fixed with cold 100% CH₃OH for 15 min, washed with PBS and blocked 1 h in 1% (w/v) BSA in PBS, 0.3% (v/v) Triton X-100. Then, cells were incubated with 1:200 lamin antibody (Santa Cruz Biotechnology) in 1.5% (w/v) BSA in PBS, 0.3% (v/v) Triton X-100 overnight at 4 °C. AlexaFluor 633 anti-rabbit IgG (Thermo Scientific, Waltham, MA, USA) was used as secondary antibody diluted 1:500 in PBS, 1% (w/v) BSA.
For phalloidin staining, cells were fixed in 3.7% (v/v) PFA in PBS for 10 min at room temperature and blocked in 1% (w/v) BSA, 0.05% (v/v) Triton X-100 in PBS for 30 min. Then, cells were incubated with Alexa 488-phalloidin (Molecular Probes, Invitrogen, Carlsbad, CA, USA) diluted 1:40 in 1% (w/v) BSA in PBS for 1 h at room temperature. Nuclei were counterstained by 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich). Samples were examined by TCS SP2-Leica confocal microscope (Leica, Wetzlar, Germany).
For phalloidin staining, cells were fixed in 3.7% (v/v) PFA in PBS for 10 min at room temperature and blocked in 1% (w/v) BSA, 0.05% (v/v) Triton X-100 in PBS for 30 min. Then, cells were incubated with Alexa 488-phalloidin (Molecular Probes, Invitrogen, Carlsbad, CA, USA) diluted 1:40 in 1% (w/v) BSA in PBS for 1 h at room temperature. Nuclei were counterstained by 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich). Samples were examined by TCS SP2-Leica confocal microscope (Leica, Wetzlar, Germany).