Fig. 7.

Tumor vessel functions, hypoxia, and antitumor effects on shKRAS CRC tumors in response to various antibody treatments. (A and B) Vessel perfusion and permeability were analyzed by injection of rhodamine-labeled lysine 2,000 kDa and 70 kDa dextran (green) in HCT116 KRAS mutant (A) and shKRAS (B) CRC tumors. Microvessels (red) are counterstained. CA9 signals represent hypoxia. (C) Quantification of vessel perfusion area and ratio, permeability, and CA9⁺ signals in HCT116 KRAS mutant and shKRAS CRC tumors (n = 10 fields per group). (D and E) Quantification of vessel perfusion area and ratio, permeability, and CA9⁺ signals in various antibody-treated HCT116 KRAS mutant (D) and shKRAS (E) tumors (n = 10 to 12 fields per group). Vehicle-treated controls are the same as those in C. (F and G) Proliferative (Ki67) and apoptotic (CC3) cell signals in various antibody-treated HCT116 KRAS mutant (F) and shKRAS (G) CRC tumors. Nuclei are counterstained with DAPI (blue). (H) Quantification of Ki67⁺ proliferative and CC3⁺ apoptotic cell signals in HCT116 KRAS mutant and shKRAS CRC tumors (n = 10 fields per group). (I and J) Quantification of Ki67⁺ proliferative and CC3⁺ apoptotic cell signals in various antibody-treated HCT116 KRAS mutant (I) and shKRAS (J) CRC tumors (n = 10 fields per group). Vehicle-treated controls are the same as those in H. All data represent mean ± SEM. (Scale bar, 50 μm.) Statistical analysis was performed using two-sided unpaired t tests (C and H) and one-way ANOVA followed by Tukey’s multiple comparison tests (D, E, I, and J). *P < 0.05, **P < 0.01, ***P < 0.001. N.S., not significant. CC3, cleaved caspase-3.