2.9. Western Blot Analysis

Proteins were obtained from Neuro-2a cells by lysis 1X using RIPA buffer containing protease inhibitor cocktail (PMSF) and quantified using Bradford assay. Briefly, protein was electrophoresed through 10% SDS polyacrylamide gel, followed by transferring to PVDF membranes and blocking (5% skim milk). The membranes were subjected to immunoblot analysis using respective antibodies (SIRT1, Nrf2, and β-actin (Supplementary Materials)). Full images of the blots are provided in the supplementary materials (Figure S2).