Western Blot Analysis of PIAS1 and Lamin A

For western blot analysis cells were washed with Phosphate-buffered saline (PBS) and lysed in Radioimmunoprecipitation assay (RIPA) buffer with complete Mini EDTA-free protease inhibitor tablets (Roche, Welwyn Garden City, UK) and the phosphatase inhibitor cocktail PhosSTOP (Roche, Welwyn Garden City, UK). The protein quantification and western blot was performed as described earlier [16 (link)]. PIAS1 XP^® Rabbit mAb (1:500, D33A7, Cell Signaling, Frankfurt am Main, Germany), Anti-β-Actin monoclonal (1:5000, AC-15, Sigma-Aldrich GmbH; Munich; Germany) and Anti-Lamin A antibody (1:1000, Abcam, Cambridge, UK) were used as primary antibody. Rabbit Anti-Mouse IgG, HRP and Goat anti-rabbit IgG, HRP (both: 1:1000, Dako, Frankfurt, Germany) were used as secondary antibody.