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. Author manuscript; available in PMC: 2012 Oct 1.
Published in final edited form as: Dev Biol. 2011 Jul 26;358(1):9–22. doi: 10.1016/j.ydbio.2011.06.045

Figure 2. Disruption of VEGF isoform expression affects normal vessel formation in developing neuroepithelium.

Figure 2

(A) Determination of relative mRNA expression was quantified by qPCR for PECAM and SSeCKs/Gravin in E9.5 neuroepithelium from wild type and VEGF120 (120/120) mice. Data are from seven embryos collected from at least three litters. Two-way ANOVA analysis of expression indicated that there was a significant source of variation between the genes (45.87%; p<0.0001) with genotype accounting for 5.73% of the variation (p=0.10); the significance from Bonferroni’s post-hoc tests are indicated in the figure. (B) VEGF protein levels in 50 μg total protein were measured in forebrain lysates from E11.5 neuroepithelium using a standard mouse ELISA assay. Data are from seven embryos collected from three litters. Both populations had a normal distribution, but unequal variance (F test, p = 0.003). While the VEGF120 mean value was 20% higher than the wild type mean, the difference was not statistically significant (Mann-Whitney test, p = 0.32). (C) Immunolabeling for Id1 (red) or NG2 (red) in combination with G. simplicifolia lectin~FITC (green) is shown for sections from E11.5 wild type and VEGF120 littermates. Id1-positive endothelial cells were present in the neuroepithelial vessels (arrows) and in the pial vessels in both wild type (left top panel) and VEGF120 (middle top panel). Lectin co-labeling highlighted the differences observed in vessel formation between wild type and VEGF120 mice. Rabbit IgG as a negative control is shown for comparison (right top panel). Scale is 25 μm in the top and bottom panels. NG2-positive pericytes process (arrowheads) with associated lectin-labeled vessels are shown in the bottom panels for wild type (left bottom panel) and VEGF120 (right bottom panel). (D) Immunolabeling for phosphohistone H3 (PHH3) as a marker of mitotic cells is shown for E13.5 dorsolateral telencephalon (Tn) in combination with lectin~FITC. The anterior loop of the forebrain is shown with arrows indicating PHH3-positive cells adjacent to investing blood vessels (green, arrowheads). The scale bar is 200μm.