Figure 3

Autophagy facilitates VCAM-1 expression via a MAPK-independent mechanism. (A,B) HUVECs were pretreated with or without 5 mmol/L 3-MA for 30 min followed by 10 ng/mL TNFα (A) or 10 ng/mL IL-1β (B) for 5 min. Phosphorylated p38, ERK1/2 or JNK and total MAPK proteins were analyzed by western blotting. (C,D) HUVECs were pretreated with 5 mmol/L 3-MA, 10 μmol/L SB202190 (SB), 10 μmol/L PD98059 (PD) or 10 μmol/L SP600125 (SP) followed by 10 ng/mL TNFα (C) or 10 ng/mL IL-1β (D) for 24 h. VCAM-1 and ICAM-1 were analyzed by western blotting. (E and F) HUVECs were pretreated with or without 10 μmol/L SB202190 for 30 min followed by 10 ng/mL IL-1β or 10 ng/mL TNFα for 24 h (E) or 6 h (F). VCAM-1 and ICAM-1 were analyzed by western blotting. Upper panel shows a representative blot and the lower panel the quantitative analysis. Each bar denotes mean ± SD of three independent experiments. *Indicates P < 0.05.