Klf10

The cells were lysed by RIPA (Solarbio, Beijing, China) with 1 mM PMSF (Solarbio, Beijing, China). Protein-extract supernatants were quantified using a BCA protein assay kit. A total of 20 μg protein was subjected to electrophoresis using SDS-PAGE and transferred to PVDF (Bio-Rad, Hercules, CA, USA). After blocking in 5% non-fat milk for 1 h at room temperature, membranes were incubated overnight at 4 °C with primary antibodies against Col2a1 (ABclonal, Wuhan, China, 1:1000), Mmp13 (Proteintech Group, Rosemont, IL, USA, 1:1000), p16 (Abcam, Cambridge, UK, 1:1000), p21 (ABclonal, Wuhan, China, 1:1000), p62 (ABclonal, Wuhan, China, 1:1000), Lc3 (Cell Signaling Technology, Danvers, MA, USA, 1:1000), Klf10 (Santa Cruz Biotechnology, Paso Robles, CA, USA, 1:1000), Bnip3 (Santa Cruz Biotechnology, Paso Robles, CA, USA, 1:1000), and β-actin (Proteintech Group, Rosemont, IL, USA, 1:5000), followed by incubation of the blots in HRP-conjugated secondary antibodies. The representative images were acquired by ECL reagents (Solarbio, Beijing, China) using an iBright 1500 imaging system (Thermo Fisher, Waltham, MA, USA).