Fig. 4

A Quantitative mass spectrometry-based profiling of cel-p (4 μM) in the presence of excess cel (8 × , 32 μM). HMGB1 protein had a high degree of credibility. B–D Target validation of HMGB1 by pull-down, WB, and IF assays in living primary neurons under normal or OGD conditions verified that 8 × cel could completely compete the binding of cel-p to HMGB1 protein. E, F CETSA results demonstrated that cel directly bound to HMGB1 protein to decrease protein degradation with increasing temperature. G High concentrations of cel or cel-p did not affect the expression of HMGB1 in primary neurons lysate or living neuronal cells within 5 h. H–K Cel had no effect on HMGB1 expression in normal primary neurons within 48 h at 0.1–0.8 μM. L–N HMGB1 levels were decreasing with time in the M and M + cel groups. Error bars represent SEM. ^#p < 0.05 vs. control group based on a one-way analysis of variance was used (n = 6). Scale bar = 25 μm