Fig. 9: Inhibition of primary tumors and metastasis in 4T1 breast tumors.

a Schematic illustration of tumor therapeutic profile. b Survival curves of 4T1 breast tumor-bearing mice. c Primary tumor growth curves of 4T1 breast tumor-bearing mice treated with Saline, Saline + RT, αCTLA-4 + RT, H@Gd-NCPs + RT and H@Gd-NCPs + RT + αCTLA-4, (n = 10 biologically independent animals, *p = 0.0465, **p = 0.0070, ***p = 0.0003). [Gd³⁺] = 30 mg kg⁻¹, [Hemin] = 12.5 mg kg⁻¹ and [αCTLA-4] = 25 μg mouse⁻¹. Treatments were performed on days 0 and 6. X-ray radiation therapy was performed 6 h after nanomedicines intravenous injection. RT 6 Gy × 2 with fractions delivered 6 days apart (black arrow). Anti-CTLA-4 antibody was treated via intravenous injection 6 h after radiation therapy (red arrow). Data were presented as mean ± SEM. d Primary excised 4T1 breast tumors weight (n = 10 biologically independent animals, **p = 0.0055, ***p = 0.0007, *p = 0.0366). e Dynamic body weights of 4T1 breast tumor-bearing mice (n = 10 biologically independent animals). f Body weight changes of individual 4T1 breast tumor-bearing mice. g Images of lung and liver fixed by Bouin’s solution. h, i Quantification of metastatic lesions of the h lungs (***p = 0.0001, *p = 0.04) and i livers (***p = 0.0009, *p = 0.0222) (n = 10 biologically independent animals). j H&E sections of lungs (scale bar = 1 mm) and livers (scale bar = 200 μm). Data (d, e, h, i) were presented as mean ± SD. Two-sided Student’s t-test was used to calculate the statistical difference between two groups. N.S. represented non-significance, and *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source data file.