Figure 4

Resveratrol is targeted to the Akt-FoxO3a signaling pathway for Nox1 and MCP-1 expression. (A, B) Cells were treated with an indicating concentrations of LY294002 (PI3K-Akt inhibitor) for 1 h prior to stimulation with LPS for 6 h. At the end of the incubation time, total mRNAs were isolated and MCP-1 (A) and Nox1 (B) mRNAs were analyzed by real-time PCR. The mRNA levels of MCP-1 and NOX1 were normalized to the levels of β-actin mRNA. (C) Cells were treated with LPS for the indicated times. After incubation with LPS, cells were lysed and the protein lysates were then analyzed by Western blotting using specific antibodies. (D) Cells were pretreated for 1 h with resveratrol and then incubated with LPS. After incubation with LPS for 30 min, cells were lysed and the protein lysates were analyzed by Western blotting using specific antibodies. (E) Cells were treated with LPS for the indicated times. After incubation with LPS, cells were lysed and the protein lysates were analyzed by Western blotting using specific anti-FoxO3a or anti-phospho FoxO3a (S²⁵³) antibodies. (F and G) Cells were pretreated with the resveratrol or LY294002, and then incubated with LPS. After incubation with LPS for 30 min, cells were lysed and the protein lysates were then analyzed by Western blotting using specific anti-FoxO3a or anti-phospho FoxO3a (S²⁵³) antibodies. (H) Signaling mechanism involved in LPS-induced Nox1 expression in macrophages and that are targeted by resveratrol.