Figure 5

PARylated HMGB1 inhibits the binding of RAGE to PS more effectively than non-PARylated HMGB1. (A) PS-precoated 96-well plates were incubated with increasing concentrations of recombinant HMGB1 (100 μg/mL, 500 μg/mL or 1 μg/mL) for 2 h at room temperature. Recombinant mouse chimeric RAGE was then added to the wells for 2 h. The plates were washed and bound RAGE was quantified by ELISA. All assays were performed in triplicate. *P < 0.05 compared with control. (B) PS-precoated 96-well plates were incubated with recombinant HMGB1, PARylated HMGB1 or BSA (1 μg/mL) for 2 h at room temperature. Recombinant mouse chimeric RAGE was then added to the wells for 2 h. The plates were washed and bound RAGE was quantified by ELISA. All assays were done in triplicate. *P < 0.05 compared with control. ^#P < 0.05 compared with HMGB1.