Figure 6 from Inhibition of Notch4 Using Novel Neutralizing Antibodies Reduces Tumor Growth in Murine Cancer Models by Targeting the Tumor Endothelium

Anti-Notch4 treatment alters vessel perfusion and angiogenesis. A and B, Assessment of vessel coverage area by CD31⁺ staining in A, B16F10 melanoma treated for 4 days with two doses (days 1 and 3) of 6-3-A6 (n = 3) and B, Py8119 tumors treated for 10 days with four doses (days 1, 3, 6, and 9) of 6-3-A6 (n = 6). Right, Representative images; left, quantification of colocalization. (Scale bar = 100 μm). C and D, Assessment of tumor perfusion in syngeneic C, B16F10 melanoma (n = 3) and D, Py8119 tumors (n = 6) using intravenously injected fluorescent tomato lectin–Alex Fluor 488. Right, Representative images costained with CD31 (red); left, quantification of colocalization. (Scale bar = 100 μm). E and F, IF staining of E, CD31⁺ tumor vessels in orthotopically implanted Py8119 tumors treated for 4 weeks with E7011 (n = 4), and F, vessel perfusion by fluorescent tomato lectin–Alexa Fluor 647 labeling with CD31 in Py8119 tumors treated for an extended duration (n = 5). Right, Representative images; left, quantification of the staining or colocalization. (Scale bar = 200 μm). Statistical analysis by the t test, plotted as average ± SEM; *, P < 0.05.