Figure 5

Confirmation of gene expression. Representative RT-PCR showing the expression of mRNAs for VEGF (A) and VEGFR-2 (B) in HRPC, HUVEC cultured alone and HRPC co-cultured with HUVEC under normoxia and hypoxia conditioned for 24 h. Densitometric analysis of mRNA expression of VEGF (C) and VEGFR-2 (D) presented as a ratio of VEGF and VEGFR-2 mRNA over 18S RNA. Representative Western blot analysis using VEGF (E) and VEGFR-2 (F) antibodies confirm the expression levels of these proteins. VEGF (homodimer, molecular mass ~ 44kDa) and VEGFR-2 (homodimer, molecular mass ~ 220 kDa) were expressed on HRPC and HUVEC cells. A β-actin antibody was used to normalize densitometer tracing values for differences in loading and the transfer efficiencies. Both of the experiments were repeated three times with similar results. Figure (G, H) bar graphs of quantitative analysis of Western blot data presenting calculated means +/- S.E. of densitometer tracings values from three independent experiments.