Figure 5 from Inhibition of Notch4 Using Novel Neutralizing Antibodies Reduces Tumor Growth in Murine Cancer Models by Targeting the Tumor Endothelium

Notch4 is expressed on tumor ECs, and treatment alters the tumor vessel transcriptional profile. A, Representative flow plots of gating for CD31⁺ cells in Py8119 and B16F10 tumors. B, Histograms comparing E7011 and human IgG binding to CD31⁺ and CD31⁻ cell populations from dissociated Py8119 and B16F10 tumors (n = 3). Left, E7011 samples were concatenated and merged into a single representative histogram. Right, Graph of concatenated MFI from Py8119 and B16F10 samples bound with E7011 or human IgG. C, Heatmaps of significantly upregulated and downregulated differentially expressed genes isolated from IP polysomes from tumor ECs treated with 6-3-A6 or control IgG (n = 3). Boxes indicate genes implicated in GPCR control of angiogenesis (red), endothelial nitric oxide regulation (blue), and inflammatory/immune response (green). D, Reactome pathway analysis of upregulated and downregulated pathways was performed using g:Profiler, and significance threshold by Benjamini–Hochberg FDR was set at 0.05. A, Fold cutoff of −log₁₀(Padj) > 2 was used to select the displayed pathways. Statistical analysis performed with the t test for perfusion studies, plotted as average ± SEM; *, P < 0.05; **, P < 0.005. Heatmap data presented as Z-scores, which integrate both fold change and significance.