Rat Mesangial Cell Culture and Cloning

The rat mesangial cell line HBZY-1 was purchased from the China Center for Type Culture Collection (Wuhan, China), and the mesangial cells were cultured and cloned as previously described.^{48 (link)}BSA, MTT, Z-VAD-fmk, Rapamycin, Bafilomycin A1 and U0126 were obtained from Sigma (St. Louis, MO, USA). The mRFP-GFP-LC3 plasmid was purchased from Genechem (Shanghai, China). The GFP-LC3 plasmid was kindly provided by Addgene (Cambridge, MA, USA). Primary antibodies for Caspase-3, PARP, LC3, Beclin-1, p62, p-c-Raf, total ERK1/2 (t-ERK1/2), p-ERK1/2, p-MEK1/2, t-MEK, Parkin and COX IV were purchased from Cell Signaling Technology (Danvers, MA, USA), and antibodies for Bax, Bcl-2, Caspase-9 and GAPDH were purchased from Abcam (Cambridge, MA, USA).