Figure 6
Effects of CORM-2 and signal molecular inhibitor in LPS-stimulated platelet.
Platelets were stimulated by LPS and treated with CORM-2 as described in Fig. 1. The platelets were lysed in RIPA buffer that contained protease and phosphatase inhibitor cocktails. Signal molecular expression and phosphorylation were assessed in LPS-stimulated platelets with treatment of CORM-2 or signal molecular inhibitors. The increase of Akt production and phosphorylation were abolished simultaneously in LPS-stimulated platelet treated with LY294002, SH-6 or CORM-2 (B,H,M). Meanwhile, GSK-3β expression and phosphorylation were inhibited in LPS-stimulated platelet with administration of LY294002, SH-6 or CORM-2 (E,K,P,S) and no differences were observed between the LY294002, SH-6 and CORM-2 (50 μM) group. Also, GSK-3β phosphorylation inhibitor (CHIR99021) were used in the further exploration. The increase of GSK-3β phosphorylation was inhibited in LPS-stimulated platelet treated with CHIR99021 (W). The similar phenomenon was observed in the CORM-2 groups with or without CHIR99021. No differences among the CHIR99021 and CORM-2 groups were observed. Similar results were also shown in CORM-2 pre-conditioning (A,G,N,D,J,Q,T,V) or CORM-2 delayed treatment group (C,I,O,F,L,R,U,X). Representative experiments are shown in (A–X). Quantitative analysis of protein expression and phosphorylation level was shown in Supplementary Figures 4 and 5.
