Anti β actin antibody

Total proteins were extracted from the cell lysates using RIPA lysis buffer supplemented with a protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA, 1:50 dilution). Protein concentrations of the extracts were measured with a BCA assay kit. For Western blot, samples were mixed with sodium dodecyl sulfate (SDS) loading buffer and separated on 8%,10%, 12%, or 15% SDS-polyacrylamide gel electrophoresis (PAGE). Proteins were electrophoretically transferred to a polyvinylidene difluoride (PVDF) membrane and blocked with 5% milk dissolved in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 1 h at room temperature. The membranes were probed overnight at 4 °C with the following primary antibodies: anti-α-synuclein antibody (Abcam, Cambridge, UK, ab138501, 1:1000 dilution); anti- p62 antibody (Abcam, ab91526, 1:1000 dilution); anti-LC3B antibody (Novus, St. Charles, MO, USA, NB100-2220, 1:1000 dilution); anti-HDAC4 antibody (Proteintech, Wuhan, China, 17449-1-AP, 1:600 dilution); anti-β-actin antibody (Antgene, Wuhan, China, ANT009, 1:1000 dilution). After incubation with horseradish peroxidase-conjugated secondary antibodies (1:1000 dilution) for 1 h at room temperature, western blots were revealed through chemiluminescence. The protein levels were quantified through densitometry using ImageJ v1.47 software.