Phospho ampkα thr172

AngII was from Sigma Aldrich (St. Louis, MO). CB13 (1-Naphthalenyl[4-(pentyloxy)-1-naphthalenyl]methanone), compound C (dosmorphin, (6-[4-(2-Piperidin-1-ylethoxy) phenyl]-3-pyridin-4-ylpyrazolo [1,5-a]pyrimidine)), JC-1 (tetraethylbenzimidazolylcarbocyanine iodide) Mitochondrial Membrane Potential assay kit and Mitochondrial Permeability Transition Pore (mPTP) assay kit were from Cayman Chemical (Ann Arbor, MI). Primary antibodies from Cell Signaling Technology (Danvers, Massachusetts) for phospho-AMPKα (Thr172) (1:1000, cat. 2535), AMPKα (1:1000, cat. 2603), phospho-LKB1 (1:1000, cat. 3482), LKB1 (1:1000, cat. 3047) and Abcam (Toronto, Ontario) connexin 43 (1:7500, cat. ab11370), CB1R (1:500, cat. ab23703), and CB2R (1:5000, cat. ab45942) were used.

The primary antibodies were purchased from Cell Signaling Technology (cleaved-Caspase 3 (Cat: 9664, 1:1000), PARP (Cat: 9542, 1:1000), AMPKα (Cat: 5831, 1:1000), Phospho-AMPKα (Thr172) (Cat: 4188, 1:1000)) and Abcam Trading (Shanghai) Company Ltd. (PLK1(Cat: ab17056, 1:1000), LC3Ⅱ (Cat: ab48394, 1:1000), P62 (Cat: ab56416, 1:1000)). The antibody against β-actin, which was used as the reference protein, was purchased from Sigma-Aldrich (Cat: A5441, 1:5000). The horseradish peroxidase- conjugated secondary antibodies Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (Cat: 115-035-003) and Goat Anti-Rabbit IgG (H+L) (Cat: 111-035-003) were purchased from Jackson ImmunoResearch Laboratories, Inc.

JC105 was synthesized in our laboratory (see below). Isosteviol was purchased from Sigma–Aldrich Co. Ltd. U0126 was purchased from Cell Signaling Technology. 3‐[4, 5‐dimethylthiazol‐2‐yl] 2, 5‐diphenyl‐tetrazolium bromide (MTT) was purchased from VWR Life Sciences. Dulbecco's modified Eagle's medium (DMEM), foetal bovine serum (FBS), 0.05% Trypsin‐EDTA (1×), penicillin (10 000 U/mL)‐streptomycin (10 000 µg/mL) (Pen Strep), phosphate‐buffered saline (PBS, 1×), no glucose and no sodium pyruvate DMEM were purchased from Gibco. RIPA buffer was obtained from Biosharp. Protease inhibitor (100x) was obtained from Beyotime. Bicinochoninic acid (BCA) assay kit was obtained from Beyotime. Stock concentration of JC105 (100 mM) and U0126 (10 mM) was dissolved in DMSO. MTT (5mg/ml) was dissolved in DMEM. During all treatments, the concentration of DMSO was below 0.1% (v/v). Phospho‐ERK 1/2 (Thr²⁰²/Tyr²⁰⁴), Total‐ERK1/2, Phospho‐DRP1 (Ser⁶¹⁶), Total‐DRP1, Phospho‐AKT (Ser⁴⁷³) and Total‐AKT were obtained from Cell Signaling Technology. Phospho‐AMPKα (Thr¹⁷²) and Total‐AMPK were obtained from Abcam and GAPDH was obtained from Proteintech. Anti‐rabbit IgG and anti‐mouse IgG were obtained from Proteintech. Enhanced chemiluminescence reagent was obtained from Affinity Biosciences.