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Figure 4 | Respiratory Research

Figure 4

From: VEGF is upregulated by hypoxia-induced mitogenic factor via the PI-3K/Akt-NF-κB signaling pathway

Figure 4

HIMF increases the transcription activities, but not mRNA stability of VEGF in MLE-12 cells. (4A) MLE-12 cells were co-transfected with pGL-VEGF and pRL-TK. Twenty-four hours later, the cells were incubated with HIMF protein as indicated. Then, cells were lysed with passive lysis buffer, and luciferase activity was measured according to the dual-luciferase assay manual. The time-course study demonstrated that HIMF-induced (40 nmol/L) VEGF transcription started at 6 h, and persisted for 24 h. After incubation with 10–80 nmol/L of HIMF, VEGF transcripts in MLE-12 were enhanced in a dose-dependent manner. (4B) MLE-12 were treated with different concentrations of HIMF and incubated with 5 μg/ml of Actinomycin D for 4, 8 and 16 h. RT-PCR indicated that HIMF did not prevent Actinomycin D-facilitated VEGF degradation in MLE-12 cells. The symbol (*) indicates a significant increase from MLE-12 controls without HIMF (P < 0.05). Triplicate experiments were performed with essentially identical results.

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