GU81-alone but not in combination with doxorubicin induces VEGF expression in vivo and in vitro. Tumor sections from each group were evaluated for VEGF expression (A&B) using immunohisochemistry. The percent VEGF positive area was calculated using Elements software and is displayed as the mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. * p < 0.05 vs control, t-test. C&D) Met-1 cells were treated with serum free DMEM +/- 2.5 uM GU81 for 24 hr, 48 hr, and 72 hr. mRNA and tumor conditioned media (TCM) was isolated and analyzed for VEGF levels at each time point. (C) Quantitative real-time PCR analysis reveals a significant increase in VEGF gene transcription following 72 hrs of GU81 treatment. * p < 0.05 vs SFM, 1-way ANOVA (D) VEGF protein levels were measured in Met-1 TCM using the Quantikine mouse VEGF ELISA kit (R&D Systems). VEGF secretion was significantly increased at all time points. ** p < 0.01, p < 0.001 vs SFM, 1-way ANOVA.