Fig. 7: Enhanced binding between LPS and HMGB1 in sera of LPS-injected MCAO animals and inhibition of LPS-HMGB1 binding by blocking HMGB1.

Binding between HMGB1 and LPS was examined by co-immunoprecipitation-immunoblotting assay. a Serum was prepared from MCAO or PSI animals at 1 or 2 days post-MCAO and precipitated with anti-LPS antibody, and amounts of HMGB1 were determined by immunoblotting using anti-HMGB1 antibody. Amounts of HMGB1, LPS, or serum albumin before immunoprecipitations are presented as input controls. b Serum was prepared from MCAO or PSI animals at 2 days post-MCAO after pre-administrating HMGB1 A box (5 μg/kg) or HPep1 (5 μg/kg) at 21 h post-MCAO, and the same co-immunoprecipitation experiments were carried out. c, d Co-immunoprecipitation-immunoblotting assay was carried out using brain tissues prepared from the cortical penumbra of MCAO or PSI animals at 2 days post-MCAO. Results are presented as mean ± SEM (n = 4). Sham sham-operated rats. **p < 0.01 vs. treatment-naive MCAO controls, ^##p < 0.01 vs. LPS-treated MCAO group