Mouse anti gcn5l1 antibody

Proteins of cells or kidney tissues were extracted by using RIPA buffer supplemented with 1% protease inhibitor and phosphatase inhibitors. After diluted in 4× SDS-PAGE loading buffer and denatured in 95°C for 10 min, samples were separated by SDS-PAGE and subsequently transferred to PVDF membranes. The membranes were blocked with 5% skimmed milk and then incubated with rabbit anti-GCN5L1 antibody (Proteintech, 19687-1-AP), mouse anti-GCN5L1 antibody (Santa Cruz, sc515444), rabbit anti-SOD2/MnSOD (acetyl K68) antibody (Abcam, ab137037), rabbit anti-MnSOD antibody (Proteintech, 24127-1-AP), rabbit anti-NLRP3 antibody (Abcam, ab210491), mouse anti-caspase-1 antibody (Santa Cruz, sc392736), rabbit anti-IL18 antibody (Proteintech, 10663-1-AP), rabbit anti-IL1beta antibody (ABclonal, A11370), rabbit anti-E-cadherin antibody (Proteintech, 20874-1-AP), and rabbit anti-αSMA antibody (Proteintech, 55135-1-AP). Then, the membranes were incubated with the corresponding secondary antibody (HRP-tagged goat anti-mouse or anti-rabbit IgG) and detected by enhanced chemiluminescence reagents (ECL, Millipore, USA). Quantitative analysis was performed using the ImageJ software.

Cells were fixed with 4% paraformaldehyde for 20 minutes, permeabilized in 0.5% Triton X-100 for 10 minutes, and then blocked with 1% goat serum for 1 hour at room temperature. Then, cells were incubated with mouse anti-GCN5L1 antibody (Santa Cruz, sc515444), rabbit anti-E-cadherin antibody (Proteintech, 20874-1-AP), and rabbit anti-αSMA antibody (Proteintech, 55135-1-AP) overnight at 4°C. Secondary antibody (Alexa Fluor 488 Goat Anti-Rabbit IgG H&L, ab150077, 1 : 500) was used to stain the cells, and DAPI nuclear stain was used to counterstain. Images were obtained using a Nikon microscope.