Fig. 3. Effect of thrombin + PC-S195A on HMGB1-mediated expression of adhesion molecules. HMGB1-mediated (1 μg/ml) expressions of VCAM-1 (white bar), ICAM-1 (gray bar), and E-selectin (black bar) at protein (ELISA, A) and transcription (mRNA; RT-PCR, B) levels in HUVECs were analyzed after pretreating cells with PC-S195A (50 nM for 30 min) and incubating them with thrombin (2 nM) for 3 h. (C) HMGB1 (1 μg/ml for 16 h)-mediated adherence of monocytes to HUVEC monolayers was analyzed after pretreating cells with PC-S195A (50 nM for 30 min) and activating them with different concentrations of thrombin for 3 h. (D) HUVECs were treated as in (C) and the HMGB1-mediated migration of monocytes through HUVEC monolayers was analyzed as described under “Materials and Methods”. Results are expressed as means ± S.D. of at least three experiments. **P ＜ 0.01 vs. HMGB1 alone.
