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. 2017 Jan 31;7:41510. doi: 10.1038/srep41510

Figure 5. Necroptosis inhibition does not prevent deferasirox-induced tubular cell death.

Figure 5

(A) Tubular cells were treated with different concentrations of Nec-1 (RIPK1 inhibitor) 1 h before addition of 10 μM deferasirox for 24 h. Nec-1 did not improve cell survival (MTT assay) at any dose. The cytokine cocktail 100 ng/ml TWEAK/30 ng/ml TNFα/30 U/ml interferon-γ (TTI) was used as a positive control for necroptosis and Nec-1. Mean ± SEM of three independent experiments. *p < 0.003 vs control. (B) Cells were pretreated with 25 μM zVAD and/or 30 μM Nec-1 before exposure to 10 μM deferasirox for 24 h. Cell death was measured by flow cytometry of annexin V/7-AAD stained cells. Mean ± SEM of three independent experiments. *p < 0.003 vs control. (C) RIPK3 expression in tubular cells transfected with a specific siRNA. Representative western blot of three independents experiments. Full-length blot is presented in Supplementary Figure 3. (D) RIPK3 was targeted with siRNA, and 48 h later cells were exposed to 10 μM deferasirox for 24 h. Cell death was assessed by flow cytometry of annexin V/7-AAD stained cells. Mean ± SEM of three independent experiments. *p < 0.003 vs control.