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. 2020 Aug 20;11(8):666. doi: 10.1038/s41419-020-02898-9

Fig. 5. The improvement of S5 in CLP sepsis was greatly dependent on autophagy.

Fig. 5

a The same samples of Fig. 2f were used to detect p62 and LC3B expression by western blot. β-actin was used as loading control. Data are means ± SEM, n = 10 mice for each experimental group. Two samples of each group were shown as representatives. *P < 0.05, **P < 0.01 vs. CLP group. b Intragastric administrated C57BL/6 mice with 20 mg/kg S5 and 50 mg/kg CQ for 2 h before suffering CLP surgery. Survival rate was observed and calculated (n = 10 mice per group). Data are means ± SEM, n = 10 mice for each experimental group. *P < 0.05, **P < 0.01 vs. CLP group. c In vitro, BMDMs were incubated with 20 μM S5 or the same volume of DMSO in the presence of 10 ng/mL LPS with 10 ng/mL IFN-γ for 6 h, or previous knockdown of Beclin1 for 36 h. Macrophage-depleted mice were treated with normal-BMDM, LPS and IFN-γ-BMDM, or S5-treated LPS and IFN-γ-BMDM and S5-treated LPS and IFN-γ-BMDM with Beclin1 knockdown (i.v.), and challenged with 10 mg/kg LPS (i.p.). Survival rate was observed and calculated. Data are means ± SEM, n = 10 mice for each experimental group. *P < 0.05, **P < 0.01 vs. M1 + BMDM group. d P62, LC3B, p-P65, P65, p-STAT1, and STAT1 expression in cells of BAFL were detected by western blot. Data are means ± SEM, n = 10 mice for each experimental group. ns nonsignificant, *P < 0.05, **P < 0.01 vs. M1 + BMDM group. e Mice were treated as Fig. 4c, 4 h later, BAFL cytokines were measured by ELISA. Data are means ± SEM, n = 10 mice for each experimental group. ns nonsignificant, *P < 0.05, **P < 0.01 vs. M1 + BMDM group.