Figure 6. HMGB1 function and Cys modification.
A. Cells expressing wild type p75 (p75wt) or mutated mitochondrial protein p75 (p75DA) were UV irradiated. Supernatants were collected for 0–6 hr or 6–24 hrs. HMGB1 was separated on a non-reducing SDS-PAGE gel and detected by western blotting with a polyclonal anti-HMGB1. B. Recombinant HMGB1 (rHMGB1), rHMGB1-C23S mutant (500ng/ml) rHMGB1-C45S (500ng/ml) mutant, or rHMGB1-C106S (500ng/ml) mutant was treated with DTT or H2O2 and added to TNP-DC that had been fed apoptotic spleen cells for 6 hrs. After overnight culture DC were harvested and injected i.v. into recipient mice that were immunized s.c. with TNBS 2 days later. Four days following immunization, mice were challenged with TNBS in the right footpad and PBS in the left footpad. Measurements (μm ± standard error) were taken 24 h later, and represent the difference between right footpad (antigen challenge) and left footpad (PBS challenge). Immune Control groups were injected with TNBS s.c. * denotes significantly different from Immune Control mice.