Fig. 2

Erbin deficiency exacerbates autophagy impairment in sepsis mice and MDP-treated BMDMs. A, C WT and Erbin^−/− mice were subjected to CLP 6 h, and the mouse liver and lung tissues were examined by transmission electron microscopy (TEM). B, D Autophagosomes were quantified by analysis of (A) and (C) results. The arrow indicates autophagosomes. Scale bar: 1 μm. E Western blotting was used to estimate the protein levels of LC3-I/II and p62 at 6 h, 24 h, and 48 h after CLP in mice liver tissue. F Western blotting was used to estimate the protein levels of LC3-I/II and p62 at 6 h, 24 h, and 48 h after CLP in mice lung tissue. G CQ (60 mg/kg) was used as a pretreatment before CLP to inhibit autophagosome-lysosome fusion. The protein expression of LC3-I/II and p62 in liver tissues were detected by western blotting. H CQ (60 mg/kg) was used as a pretreatment before CLP to inhibit autophagosome-lysosome fusion. The protein expression of LC3-I/II and p62 in lung tissues was detected by western blotting. I BMDMs were examined by transmission electron microscopy (TEM). The arrow indicates autophagosome. Scale bar: 1 μm. J Autophagosomes were quantified by analysis of results of (I). K The LC3-I/II fluorescence was detected by confocal microscopy, scale bar: 50μm. L The LC3-I/II fluorescence staining was quantified by analysis of results of (K). M Erbin^fl/fl and Erbin^{fl/fl/Lyz2−cre} BMDMs were treated with MDP (6 h), and the western blotting was used to estimate the protein levels of LC3-I/II, p62, Beclin1, ATG5, and ATG7. 200 nM RAPA was used as a positive control for autophagy. N CQ (5 μM) was used as a pretreatment before MDP to inhibit autophagosome-lysosome fusion. The LC3-I/II conversion and p62 accumulation were detected by western blotting in BMDMs. *P < 0.05, **P < 0.01, ***P < 0.001 from ANOVA followed by Tukey’s post hoc test