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. 2013 Sep 25;5:17. doi: 10.1186/2045-824X-5-17

Figure 5.

Figure 5

Combined Notch and VEGF decreases endothelial cell coverage. (A) Representative pictures of PECAM-1 immunofluorescence (green) of NGP-LacZ controls (upper left), NGP-N1D (upper right), NGP-LacZ + BV (lower left), and NGP-N1D + BV (lower right). Bar = 100 μm. (B) Quantification of PECAM-1 immunofluorescence. NGP-N1D and NGP-LacZ + BV tumors had decreased PECAM-1 than NGP-LacZ controls. NGP-N1D + BV tumors had an additive decrease in PECAM-1. Error bars represent SD. **P < 0.01, ***P < 0.001. (C) Representative pictures of triple staining for TUNEL (red), PECAM-1 (green), DAPI (blue), white arrowheads. Bar = 25 μm (D) The number of DAPI(+), TUNEL(+) cells surrounded by PECAM-1 in each tumor normalized by the viable area shows increased apoptotic ECs in NGP-N1D + BV tumors. Error bars represent SD. **P < 0.01. (E) HUVECs incubated with N1D and BV had increased apoptosis compared to HUVECs with BV only. HUVECs incubated with N1D and VEGF had decreased apoptosis compared to N1D with BV, but were not different from HUVEC incubated with BV only. Consistent with the known role of VEGF promoting endothelial cell survival, HUVEC incubated with VEGF had lower apoptotic levels than HUVECs incubated with BV, and lower apoptotic levels than HUVECs incubated with N1D and VEGF. Error bars represent SD. *P < 0.05. (F) N1D with BV resulted in increased BrdU incorporation by HUVECs compared to BV only. Addition of VEGF did not affect N1D-induced proliferation. Error bars represent SD. *P < 0.05.