Fig. 1: Saracatinib inhibited necroptosis in different cell lines.

A L929 cells were pretreated with individual compound (10 μM) for 1 h following treatment with TNF (10 ng/ml), Smac mimetic (SM-164) (0.1 μM) and pan-caspase inhibitor z-VAD (10 μM) (TSZ) to induce necroptosis. Cell viabilities were determined with Cell Counting Kit-8 (CCK8) method and normalized to control (DMSO only). NEC-1 (20 μM) and GSK872 (10 μM) were chose as positive control. The screen data for each compound was singlicate. B Chemical structure of saracatinib cited from TargetMol. C L929 cells were pretreated with saracatinib at different concentrations for 1 h prior to treatment with TSZ. Cell viabilities were determined by CCK8 method (n = 3). D L929 cells were treated with saracatinib at different concentrations for 24 h. Then cell viabilities were determined by CCK8 method (n = 6). E NIH-3T3-RIPK3 cells, Hela-RIPK3 cells, and HT-29 cells were pretreated with NEC-1 (20 μM), GSK872 (10 μM) or saracatinib (7.5 μM) for 1 h prior to treatment with TSZ. Cell viabilities were determined by CCK8 method (n = 6 for NIH-3T3-RIPK3 cells, n = 3 for Hela-RIPK3 cells, n = 8 for HT-29 cells). F Bone marrow-derived macrophages (BMDM) were pretreated with NEC-1 (20 μM), GSK872 (10 μM) or saracatinib (15 μM) for 1 h prior to treatment with LPS + z-VAD. Cell viabilities were determined by CellTiter-Glo Luminescent Cell Viability Assay (n = 5). G L929-RIPK1 KO cells were seeded in 24-well plates and pretreated with NEC-1 (20 μM), GSK872 (10 μM) or saracatinib (15 μM) for 1 h prior to treatment with IFN-γ (20 ng/ul) and z-VAD (10 μM). Cells were harvested and incubated with propidium iodide (PI) (n = 3). PI negative cells were analyzed by flow cytometry. **: p < 0.01. Data are represented as mean ± SD. See also Fig. S1.