Fig. 5.

Nec-1 prevents caspase activation in cultured tubular cells. (A) Cleaved caspase-8 and cleaved caspase-3 proteins in tubular cells exposed to TTI or TTI + zVAD for 6 h following pretreatment with Nec-1. (B) Cleaved caspase-3 staining, detected by immunofluorescence. Cleaved caspase 3 is shown in green and DAPI, in blue. Original magnification, 630×. (Scale bars: 15 µm.) (C) MMP in tubular cells at 24 h of treatment. Graph shows TMRM staining. Data are expressed as mean ± SEM of three independent experiments. ***P < 0.001 vs. control; ^###P < 0.001 vs. TTI or TTI/zVAD vehicle. (D) Cleaved caspase-8 and cleaved caspase-3 proteins in tubular cells transfected with a RIPK1 or control siRNA and exposed to TTI for 6 h. (A and D) Representative Western blots of three independent experiments are demonstrated.