Figure 4 | Laboratory Investigation

Figure 4

From: Antagonism of Ang-Tie2 and Dll4-Notch signaling has opposing effects on tumor endothelial cell proliferation, evidenced by a new flow cytometry method

Figure 4Figure 4

BrdUrd uptake analysis of TAECs and GFP-expressing tumor cells in U-87 human tumor xenografts. (a, b) Flow cytometry analysis, dual parameter dot plots representing tumor xenograft cells stained with anti-CD31, anti-CD45, and anti-BrdUrd antibodies. (a) Gated regions were set on GFP-positive (green, human tumor cells) and GFP-negative (black, mouse cells) cell populations. Additional gated region set on the CD31high/CD45neg cell population (red). (b) BrdUrd uptake for TAECs and U-87-GFP tumor cells (n=3) were determined with respective percentage of BrdUrd (+) population ranges of 20–29% (lower panel) and 33–38% (upper panel). (c) IHC analysis of U-87-GFP tumor co-labeled with anti-CD31 and anti-BrdUrd antibodies. CD31-labeled microvascular structures (blue) and BrdUrd-labeled nuclei (brown). Image gallery of representative BrdUrd-labeled CD31 nuclei (red arrows) with distinct nuclear morphology compared with BrdUrd-labeled tumor nuclei (lower panel). Scale bar=100 μm (upper panel), enlargements of representative views are shown in lower panel.

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