Figure 1.

qRT-PCR analysis of the effect of VEGF/anti-VEGF on expression of Wnt pathway members. Triplicate dishes of HG HRECs were treated with either VEGF vehicle (black bar), or VEGF (1 nM; red bar) for the indicated duration and then harvested. The green bar indicates the response of cells that were treated with VEGF for 8 hours and then anti-VEGF for an additional 8 hours. Cells were lysed, RNA was extracted and subjected to qRT-PCR using specific primers. The level of expression in unstimulated cells (black bars) was set to 1.0. The data are expressed as the mean ± SD (n = 3–4). For VEGF-treated cells (red bars) the data are expressed as fold increase over unstimulated cells, *P < 0.05. The green bar shows the response of cells that were treated with VEGF for eight hours and then anti-VEGF for an additional 8 hours; statistically significant differences between these two experimental conditions are indicated by #P < 0.05. RNAseq and qRT-PCR analysis indicated that VEGF-induced change in expression persisted to 24 or 48 if anti-VEGF was not added (reference⁷ and Supplementary Fig. S1). Similar results we observed in three independent experiments.