FIGURE 2.

LPS-induced CAM levels in HUVECs are downregulated by 6-shogaol in a concentration-dependent manner. Confluent HUVECs were treated with indicated concentrations of 6-shogaol. After 30 min, the cells were activated by LPS (1 μg/ml) for (A,B) 4 h (E-selectin) or 24 h (ICAM-1 and VCAM-1) to determine protein levels and for (C) 2 h (E-selectin) or 12 h (ICAM-1 and VCAM-1) for mRNA expression. (A) Surface protein levels of ICAM-1, VCAM-1, and E-selectin were determined by flow cytometry, and (B) total protein levels of CAMs were analyzed by Western blot analysis. Actin served as the loading control. One representative blot is shown. (C) mRNA data were obtained by qPCR. GAPDH served as the housekeeping gene. 6-SG: 6-shogaol. Data are expressed as mean ± SD; (A,C) n = 3; (B) n = 4/4/3; *p ≤ 0.05 vs. LPS.