Figure 5
FGF-2 induces VEGF mRNA and protein in Balb/c 3T3 embryonic fibroblasts. A, Serum-restricted fibroblasts were treated with FGF-2 (10 ng/ml) (F), hypoxia (2% O2) (H), or untreated control (C). Northern blot analysis for VEGF (VEGF) and ribosome-associated protein (36B4) was performed (exposure time: 24 hours). B, VEGF protein secretion from control (−FGF-2) or FGF-2–treated (+FGF-2) Balb/c 3T3 fibroblasts over an 8-hour time period. Triplicate plate conditioned media samples were analyzed by ELISA and represented as average VEGF (ng/ml) ± sem. Statistically significant difference from untreated samples (* p < 0.05). C, Transient transfection and transcriptional activation of VEGF-promoter-luciferase reporter construct. Balb/c 3T3 cells were transfected in triplicate with either the VEGF promoter-luciferase construct or the pGL3 control vector. Postrecovery (18 hours), cells were treated with FGF-2 (FGF-2), hypoxia, or nothing (Control) for 8 hours. Cell lysates were analyzed for luciferase activity and subtracted from the pGL3 control levels. Data is presented as average light units per mg protein ± sem.
