Figure 1.

CA4-NPs reduced the tumor burden. H22 cells were subcutaneously inoculated into Balb/c mice. When the tumor volume reached ~170 mm³, CA4-NPs were intravenously administered at 45 mg/kg (on a CA4 basis) on day 0. (A) Schematic image showing the preparation of CA4-NPs. (B) IHC staining of CD31 in the tumors on day 2 after CA4-NP treatment. (C) Quantitative analysis of the microvessel density (MVD, which is represented as No. per field) for the IHC staining of CD31 in tumors. Seven high-density vascular areas (“hot spots”) of tumors were observed using an optical microscope at 100×, and then CD31⁺ vessels were photographed and counted at 400× in each respective hot spot. (D) IHC staining of Ki67 in the tumors on day 2 after CA4-NP treatment. (E) Quantitative analysis of the Ki67⁺ area (Ki67⁺ area/field) in IHC staining using Image J software (n = 5). (F) H&E staining of tumors in the PBS and CA4-NP groups on day 2. V, viable region of tumors; N, necrotic region of tumors. (G) Quantitative analysis of the necrotic area (necrotic area/field) for H&E staining of the tumors using Image J software (n = 3). (H) Tumor burden (long axis of tumors) in the PBS and CA4-NP groups on day 8 (n = 5). (I) ELISA for evaluating the intratumoral VEGF level at 0 h, 24 h, 48 h, and 72 h post-CA4-NP treatment (n = 5). Data are presented as mean ± SD (**P < 0.01, ***P < 0.001, ****P < 0.0001, ns, not significant).