Acetylation of HMGB1 by JNK1 Signaling Promotes LPS-Induced Peritoneal Mesothelial Cells Apoptosis
Influence of Wt-HMGB1 and mut-HMGB1 on LPS-induced HMrSV5 cells apoptosis. (a) Apoptosis in LPS-stimulated HMrSV5 cells was assessed by flow cytometry. (b) The apoptotic rate among different groups. Data are means±SE (n=3). p<0.05 versus LPS-untreated cells. (c) HMrSV5 cells stably expressing Wt-HMGB1 or mut-HMGB1 were exposure to LPS (5ug/ml) for 48 hours. The empty vector pEGFP-N1 was used as a mock-transfection control. Exogenous and endogenous acetylated HMGB1 were examined by coimmunoprecipitation. (d) The acetylation levels of endogenous and exogenous HMGB1 were quantitatively analyzed by densitometer. (e) DNA contents were analyzed by flow cytometry. (f) The ratio of cells at sub-G1 stage. (g) The levels of BAX and cleaved-caspase3 were examined by immunoblotting assay. (h) Expression levels of the indicated proteins were quantitatively analyzed by densitometer and normalized with β-actin. Data in (d), (f) and (h) are means±SE (n=3). p<0.05 versus same group without LPS exposure; †p<0.05 versus Wt-HMGB1 transfected group with LPS treatment.
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