Fig. 6

CAs’ pharmacological inhibition increased vessel maturity and decreased lung metastasis. a Tumor microvessel density (MVD) was analyzed by quantifying the CD31-positively stained area in tumor sections from each group by ImageJ. Blood vessel hotspots were selected (25 hotspots per treatment group). Representative images of tumor sections fixed and stained with anti-CD31 antibody (red) to identify the blood vessels and counterstained with DAPI (blue). Scale bar, 50 μm. b The microvessel pericyte coverage index (MPI) was analyzed by counting the vessels which stained positively for both CD31 and α-SMA (yellow arrowheads) among all CD31-positive vessels (yellow arrows) in blood vessel hotspots (15 hotspots per treatment group). Scale bar, 50 μm. c Images of whole tumors resected from the mice under each treatment condition after 27 days of drug treatment. d Tumor volume was calculated from the measurement obtained from the tumor-bearing mice using a pair of calipers. The measurements were taken on the indicated days. e Tumor metastasis was observed by detecting tumor cell luminescence intensity in the lungs (arrowhead) using IVIS spectrum. (F) Tumor hypoxia was measured by staining Glut1 expression in the tumor sections. The Glut1 staining was observed in the tumor cells. Scale bar, 100 μm. All data presented as mean ± SD, *P < 0.01, **P < 0.001 by ANOVA