Fig. 2: Stimulation of complement triggers podocyte pyroptosis.

Podocytes were treated with or without C3a (1 μM) or C5a (100 nM). The pyroptotic features of podocytes, including GSDMD cleavage, membrane integrity, LDH release, and PI uptake, were assessed. A The expression of pyroptosis-related mRNAs was measured by qRT–PCR. B, C The levels of pyroptosis-related proteins were determined by western blotting. D The cell membrane integrity and apoptosis of podocytes were assessed using a microplate reader. E C3a and C5a promoted the release rate of LDH. F, G Microscopic imaging of podocytes stained with Hoechst 33342 and propidium iodide (PI) (scale bar = 50 μm). The right panel represents the percentage of PI-positive podocytes (6 and 16 high-power fields were analyzed in group NC and group C3a respectively in (F); 8 and 11 high-power fields were analyzed in group NC and group C5a respectively in (G)). The data in (A), (D), and (E) represent the mean ± SEM of three independent experiments. *p < 0.05.