Fig. 5

GSK872 suppressed RIP3-mediated necroptosis and restored Kir4.1 protein expression in mice within 3 days after LPS injection. A The experimental protocol. Mice were treated with GSK872 (2 mg/kg, i.p.) or vehicle (0.25% DMSO) 1 h before 4 mg/kg LPS was given. The hippocampus was obtained to assess the protein levels at 6, 48, and 72 h after LPS injection (n = 3 per group, each time point). The expression levels of p-RIP3, RIP3, p-MLKL, and MLKL detected RIP3-mediated necroptosis. B A representative Western blot images showing the specific bands for these proteins and Kir4.1 protein. An equal amount of protein sample (20 μg) obtained from the hippocampus homogenate was applied to each lane, and β-actin protein was used as the internal control. C–E Bar graphs showing the densitometric analysis of the molecules p-RIP3 and total RIP3 (C), p-MLKL and total MLKL (D), and Kir4.1 (E), normalized to total RIP3, total MLKL or β-actin expression, respectively. Each bar represents the mean ± SEM. One-way ANOVA; Bonferroni post hoc test vs. saline- and vehicle-treated groups or LPS- and vehicle-treated groups; *p < 0.05, **p < 0.01, and ***p < 0.001