Anti lc3

G‐Rb3 (purity ≥ 98.5%, HPLC method) was isolated and purified from the leaves of Panax quinquefolium (American ginseng). Cisplatin was purchased from Sigma Chemicals with purity more than 99%. Compound C, rapamycin (Ram) and acetylcysteine (NAC) also were purchased from MedChemExpress Biotech and stored at −80°C in darkness. The commercial assay kits for determining reduced glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), blood urea nitrogen (BUN) and creatinine (CRE) were bought from Nanjing Jiancheng Biological Research Institute. Haematoxylin and eosin (H&E) dying kit and Hoechst 33258 staining kit were obtained from Beyotime Co, Ltd. The immunohistochemically assay kits together with SABC‐DyLight488 immunofluorescence staining kits were obtained from BOSTER Biological Technology Co, Ltd. The primary rabbit monoclonal antibodies including anti‐LC3, anti‐BNIP3, anti‐β‐actin, anti‐GAPDH, anti‐Atg3, anti‐Atg5, anti‐Atg7 and anti‐p62 were all provided by BOSTER Biological Technology Co, Ltd. The rabbit anti‐AMPK, rabbit anti‐mTOR, rabbit anti‐Bax, Bcl‐2, Bad, caspase 3 and caspase 9 were acquired from Cell Signaling Technology. TUNEL commercial kit was purchased from Roche Applied Science. All other reagents and chemicals, unless indicated, were obtained from Beijing Chemical Factory.

Immunohistochemistry was performed with specific antibodies for target proteins following a protocol described previously with some modifications [6 (link)]. Knee joint sections on slides were incubated with anti-VDR, anti-PPAR-γ, anti-LC-3, or anti-P62 (Boster, Wuhan, China) antibodies. Subsequently, the sections were stained with a polymer horseradish peroxidase (HRP) detection system (PV9001, ZSGB-BIO, Beijing, China) and visualized with a diaminobenzidine (DAB) peroxidase substrate kit (ZLI-9017, ZSGB-BIO, Beijing, China). Each section was evaluated under a microscope (DMi8, LEICA, Wetzlar, Germany) in three randomly selected areas at a magnification of 20×. Image-Pro Plus 6 (Media Cybernetics, Inc.) was used to analyze the average integrated optical density (IOD) according to a previously described protocol [35 (link)].