Figure 3
From: VEGF is upregulated by hypoxia-induced mitogenic factor via the PI-3K/Akt-NF-κB signaling pathway
Generation of HIMF overexpressing cells. MLE-12 cells were transfected with HIMF cDNA or control vector. Stable cell lines, MLE-HIMF, along with their transfection control cells MLE-Zeo, were screened based on resistance to Zeocin (400 μg/ml). Western blots with cell culture medium for HIMF and protein from cell lysate for VEGF (3A) and RT-PCR with cell total RNA (3B) demonstrated that MLE-HIMF have higher HIMF protein and mRNA levels than their parent (MLE-12) and transfection (MLE-Zeo) counterparts. The VEGF levels in MLE-HIMF were also increased significantly compared with those of their controls. The symbol (*) indicates a significant increase from parent controls (P < 0.05). Triplicate experiments were performed with essentially identical results.