Immortalized Mouse Podocyte Injury

The immortalized mouse podocyte line MPC5 was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). To induce differentiation, MPC5 podocytes were cultured in RPMI-1640 medium (Gibco, New York, NY, USA) supplemented with 10% FBS (Gibco, New York, NY, USA) without IFN-γ for 2 weeks in a 5% CO₂ incubator (Liu et al., 2016a (link)). Differentiated MPC5 cells were used in subsequent experiments. MPC5 cells were treated with HG (30 mM) to establish an in vitro DN model, and 5 mM glucose was used as a control (low glucose). To examine the effects of Bre, MCC950, nigericin, and Ac-YVAD-cmk on MPC5 cell injury, the cells were treated with 50 µM or 100 µM Bre, 10 µM MCC950 (Merck, Rahway, MA, USA), 10 µM nigericin (MCE, Princeton, NJ, USA), and 40 µM Ac-YVAD-cmk (Merck, Rahway, MA, USA), and then cell injury was assessed.