Protein Extraction and Western Blot Analysis

The cells after the corresponding treatments were harvested and lysed with lysis buffer (20 mM Tris (pH7.5), 150 mM NaCl, 1 % Triton X-100, sodium pyrophosphate, β-glycerophosphate, EDTA, Na3VO4, leupeptin, and 1 % protease inhibitor cocktail (Roche)). 10 % or 12 % SDS-PAGE was used to separate the collected cell lysates according to the methods described previously [12 (link), 48 (link)].
Blots were probed with the specific antibodies against PCNA (Proteintech, Chicago, USA), β-actin (Proteintech, Chicago, USA), Lamin A/C (Proteintech, Chicago, USA), Tubulin (Proteintech, Chicago, USA), atp A1 (Cell Signaling Technology, USA). Secondary antibodies conjugated with Horseradish peroxidase (ProteinTech Group) and enhanced chemiluminescence (ECL kit, Beyotime, China) were employed to test the expression of proteins.