Fig. 2.

Antiangiogenic and vascular-modulatory effects of Ang2/VEGFA blockade in combination with agonist CD40 antibodies. (A) Representative images of CD31 immunostaining (red) and DAPI nuclear staining (blue) of MC38 tumors treated with the indicated antibodies. (Scale bar, 100 μm.) (B–D) Relative CD31⁺ area in (B) MC38 and (C) MMTV-PyMT and (D) flow cytometry analysis of CD31⁺ cells in B16-OVA tumors. Each data point indicates one tumor per mouse and represents an average of at least 4 images per tumor (B and C). (E) Analysis (Left) and representative image (Right) of CD31⁺ vessels in MC38 tumors treated as indicated. Vascular leakage was identified by FITC-dextran in the abluminal compartment of CD31⁺ blood vessels (arrowhead). Each data point represents one image; at least 3 representative images were analyzed per tumor. (Scale bar, 100 μm.) (F) Analysis (Left) and representative image (Right) of CD31⁺ vessels in MC38 tumors treated as indicated. Nonperfused CD31⁺ vessels were identified by absence of FITC-lectin staining (asterisks). Each data point represents one image; at least 3 representative images were analyzed per tumor. (Scale bar, 100 μm.) (G) Relative proportion of NG2⁺ pericyte-covered blood vessels in MC38 tumors treated as indicated. Each dot indicates one tumor. Data indicate mean values ± SEM. Statistical analyses by 1-way ANOVA with Tukey’s correction for multiple comparisons, unless otherwise indicated in Dataset S2. The number of mice employed in each experiment is reported in Dataset S2.