Acetylated sod2 ac sod2

The livers of septic mice were collected and lysed with RIPA buffer. Total and nuclear proteins were extracted using the RIPA Lysis Buffer (Beyotime, Beijing, China) and a nuclear extraction kit (Beyotime, Beijing, China), respectively. Protein samples were separated using 10–12% SDS-PAGE and transferred onto PVDF membranes. Then, the membranes were washed and blocked for 120 min, followed by incubation with primary antibodies against NF-κB p-p65 (Abcam, USA), histone H3 (Affinity, China), Sirt3 (Abcam, USA), PGC-1α (Abcam, USA), SOD2 (Abcam, USA), acetylated SOD2 (Ac-SOD2) (Abcam, USA), caspase-1 (Abcam, USA), GSDMD (Abcam, USA), IL-1α (Abcam, USA), IL-1β (Abcam, USA), or GAPDH (Affinity, China) at 4°C overnight. All antibodies were diluted according to the manufacturers' instructions. The membranes were washed and incubated with secondary antibodies at room temperature for 1 h. Finally, the blots were detected using enhanced chemiluminescence substrate (ECL kit, Millipore). Signal intensities were measured using Fusion software.

All chemicals and reagents (unless otherwise stated) were of analytical grade and were purchased from Sigma-Aldrich^® (Merck KGaA, Darmstadt, Germany). Antibodies against AMP-activated protein kinase (AMPK), sirtuin 3 (SIRT3), superoxide dismutase 2 (SOD2), B-cell lymphoma 2 (Bcl-2), pro-caspase3, cleaved-caspase3, phospho-dynamin-like protein 1 at Ser616 (p-Drp1^Ser616), mitofusin 2 (Mfn2), phospho-IκBα (p-IκBα), phospho-NFκB p65 (p-NFκB p65), tumor necrosis factor-alpha (TNF-α), and β-actin were purchased from Cell Signaling Technology (Danvers, MA, USA). Peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α), phospho-AMPK at Thr172 (p-AMPK^Thr172), and interleukin-1beta (IL-1β) were obtained from Merck (Merck KGaA, Damstadt, Germany). Total OXPHOS antibody cocktail, acetylated SOD2 (Ac-SOD2), and Bcl-2-associated X protein (Bax) were purchased from Abcam (Cambridge, MA, USA).