Figure 7
From: VEGF is upregulated by hypoxia-induced mitogenic factor via the PI-3K/Akt-NF-κB signaling pathway
Activation of NF-κB is essential for HIMF-induced VEGF expression. Cells were co-transfected with pNFκB-luc, dominant-negative mutants of NF-κB pathway and pRL-TK, with or without stimulation of HIMF protein for various periods as indicated. (7A) Western blots indicated that HIMF (40 nmol/L) induced phosphorylation of IKK and IκBα in MLE-12 cells. (7B and 7C) Transfection of MLE-12 cells with dominant-negative mutants IKKα (K44A) and IKKβ (K44A), and super-repressor IκBα (S32A/S36A), abolished HIMF (40 nmol/L)-induced NF-κB activity and upregulation of VEGF in MLE-12 cells. The symbol (*) indicates a significant increase from MLE-12 controls untreated with HIMF (P < 0.05). The symbol (#) indicates a significant decrease from MLE-12 cells treated with HIMF only (P < 0.05). Triplicate experiments were performed with essentially identical results.