Supplementary Figure 7: ROS and calcium are both required for necroptosis in U937 cells.
From: Plasma membrane translocation of trimerized MLKL protein is required for TNF-induced necroptosis
(a) U937 cells were either cultured in calcium free medium or cultured in calcium free medium and pre-treated with NAC (5 mM). Then, the cells were treated with TSZ. After 10 hours, cell survival was determined by PI staining. Results shown are averages of triplicates ± SEM. (b) U937 cells cultured in calcium free DMEM were treated with TSZ as indicated. Cells were harvested and the Fluo4 fluorescent cells were determined by FACS analysis using Fluo4. (c) Immunoblot analysis of U937 clones expressing control-shRNA or MLKL-shRNA with anti-MLKL or anti-Actin antibodies. * indicates non-specific band. **,P<0.01. Statistics source data for Supplementary Fig. 7 can be found in Supplementary Table 1. Uncropped images of western blots are shown in Supplementary Fig. 8.
