Fig. 5

Activation of AMPK/PKC axis promoted SR-A1 transfer to cytomembrane for engulfment of HMGB1. A Time course of SR-A1 (green) location in BMDMs treated with AICAR (300 μM). Confocal microscopy images were captured at 0, 15, 30, and 60 min after AICAR administration. Scale bar: 10 μm. B The expression of SR-A1 on the membrane of BMDMs treated with AICAR (300 μM, 15 min) (n = 3). C Phosphorylation of PKC in BMDMs treated with AICAR (300 μM, 15 min) (n = 3). D rHMGB1 (his tag) contents in BMDMs treated with AICAR (300 μM) in the presence of PKC inhibitor (Go6983, 100 nM) (n = 3). E Phosphorylation of AMPK and PKC in BMDMs treated with metformin (2.5 mM, 15 min). F rHMGB1 (his tag) contents in BMDMs treated with AICAR (300 μM) in the presence of the AMPK inhibitor (CC, 20 μM) or PKC inhibitor (Go6983, 100 nM) (n = 3). BMDMs were administered with CQ (20 μM) for 15 min, then administered with inhibitors Go6983 (D) (F) or CC (F), then treated with AMPK agonist (AICAR or metformin) for 15 min, followed by co-culturing with rHMGB1 (his tag) (10 nM) for 1 h, and subsequently collected for western blotting. Representative bands and a data summary (n = 3 for each group) are shown. A significant difference was revealed following unpaired Student’s t-test (B and C) or one-way ANOVA (D, E and F) (*p < 0.05, **p < 0.01 and ***p < 0.001 vs. Control; ^#p < 0.05, ^##p < 0.01 and ^###p < 0.001 vs. AICAR group or metformin group; Bonferroni post hoc tests)