*2.4. Protein Extraction and Quantification*

After treatment, cells were harvested, washed three times with PBS, and lysed using a cold RIPA lysis buffer supplemented with a protease inhibitor and an EDTA solution (Thermo, Hudson, NH, USA) at a ratio of 100:1:1, then centrifuged at 13,000 rpm and 4 ◦C for 30 min. The supernatant was collected, and the protein concentration was estimated with a BCA Protein Assay Kit (Sigma, St. Louis, MO, USA) using BSA as the standard.
