Figure 2. VEGF induces annexin A2 mRNA and protein expression in a time-dependent and dosage-dependent manner in RF/6A cells. A: RF/6A cells were treated with 25 ng/ml VEGF for the indicated length of time. Results of quantitative real-time PCR are representative of three independent experiments, each performed with duplicate samples. B: The protein expression of Annexin A2 was analyzed by western blot in RF/6A cells treated with 25 ng/ml. VEGF for the indicated length of time. Total cell lysates and acetone-precipitated medium from cells were analyzed for the presence of annexin A2 protein. Annexin A2 is indicated (arrows). The same membrane was blotted with anti–total Akt antibody for normalization. C: VEGF induced an increase in annexin A2 mRNA levels measured by quantitative real-time PCR in a concentration-dependent manner. RF/6A cells were treated with the indicated concentrations of VEGF for 2 h, and then total RNA was isolated.