Fig. 6. PSMα1-induced neutrophil necroptosis was blocked by WRW4 or anti-TNFα.

a, c Neutrophil were treated with PBS or TNFα neutralizing antibody at indicated concentrations for 1 h prior to the incubation with 1 μM PSMα1 for 1 h. The release of LDH was detected (n = 6) and the level of pMLKL, MLKL and β-actin in neutrophil were detected by western blot (n = 3). b, d, e Neutrophil were treated with PBS or WRW4 at indicated concentrations for 1 h prior to the incubation with 1 μM PSMα1 for 1 h. The release of LDH was detected (n = 6), the level of pMLKL, MLKL and β-actin in neutrophil were detected by western blot (n = 3), and the level of TNFα was detected by ELISA assay (n = 8). Data were shown as mean ± SE (a, b, e). *P < 0.05, **P < 0.01, ***P < 0.001 vs. control group; ^###P < 0.0001 vs. PBS-treated group