High mobility group box-1 release from H2O2-injured hepatocytes due to sirt1 functional inhibition

Figure 5 Sirt1 inhibits high mobility group box-1 translocation and release. A: Acetylated HMGB1 levels determined by immunoprecipitation. EX527, a Sirt1 inhibitor inhibited sirt1 level and promoted HMGB1 acetylation; B: High mobility group box-1 (HMGB1) expression in the nucleus or cytoplasm determined by Western blot; C: The mRNA and protein levels of Sirt1. BNL CL₂ cells were transfected with control lentiviral vector or Sirt1 shRNA lentiviral vector for 48 h. The mRNA and protein levels of Sirt1 were detected by real-time PCR and Western blot. Multiplicity of infection, refers to the number of lent virus adsorption on the cells and the ratio of the number of cells in culture; D: HMGB1 translocation; E: HMGB1 content in medium determined by ELISA. All data are presented as the mean ± SD. Statistical analysis was done by the Student’s t-test. ^bP < 0.01 vs 0 group or negative control, ^fP < 0.01 vs control lent viral group. MOI: Multiplicity of infection; HMGB1: High mobility group box-1.