Western Blot Assay Protocol for Diverse Proteins

Western blot assays were performed as described previously^{18 (link)}. Due to the diversity of the target proteins to be tested, the membrane was usually cropped into 2–4 parts based on the molecular weight of the target proteins and then the corresponding antibody hybridization was performed. The primary antibodies used in this study included p-PERK (1:1000, Abclonal, AP0886), PERK (1:1000, Cell Signaling Technology, 2683), p-NRF2 (1:1000, Affinity, DF7519), NRF2 (1:1000, Cell Signaling Technology, 12721), p-STAT3 (1:1000, Cell Signaling Technology, 9139), STAT3 (1:1000, Cell Signaling Technology, 9145), p-c-Jun (1:1000, Cell Signaling Technology, 3270), c-Jun (1:1000, Cell Signaling Technology, 9165), MyoD (1:200, Santa Cruz Biotechnology, sc-377460), connexin 43 (1:2000, Proteintech, 26980-1-AP), CYP11A1 (1:1000, Cell Signaling Technology, 14217), CYP19A1 (1:1000, Abcam, ab18995), StAR (1:1000, Cell Signaling Technology, 8449s), and β-actin (1:5000, Proteintech, 20536-1-AP). The data were obtained from triplicates of each independent experiment. Raw data for each blot were shown in the Supplementary file.