Fig. 7: Blockade of ENO1 significantly enhanced the response to radiotherapy and triggered antitumor immunity in a TNBC animal model.

A A total of 5 × 10⁵ CT26 cells were subcutaneously injected into the left legs of BALB/c mice for 5 days and then intraperitoneally administered with anti-ENO1 antibodies (HuL001, 40 mg/kg) or clodronate liposomes (50 μL/mouse) on the indicated days (n = 5). Local radiotherapy was given on Day 10. The tumor volume was recorded every three days. *p < 0.05. Two-Way ANOVA test (n = 4). B The densities of M1 (CD80⁺) and M2 (CD206⁺) macrophages in resected tumors were analyzed by immunofluorescence staining. The quantification of M1 (CD80⁺) and M2 (CD206⁺) macrophages in resected tumors. *p < 0.05 and ***p < 0.001. One-Way ANOVA test (n = 3). C 4T1 cells (5 ×10⁴) were subcutaneously injected into the left legs of BALB/c mice for 4 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 20 mg/kg) six times on the indicated days (n = 5). Local radiotherapy was given on Days 10 and 12. The tumor volume was recorded every three days. *p < 0.05 and ***p < 0.001. Two-Way ANOVA test (n = 4). D The resected tumors were weighed on Day 40. *p < 0.05. One-Way ANOVA test (n = 4). E The densities of M1 (CD80⁺) and M2 (CD206⁺) macrophages in resected tumors were analyzed by immunofluorescence staining. F The quantification of M1 (CD80⁺) and M2 (CD206⁺) macrophages in resected tumors. *p < 0.05. One-Way ANOVA test (n = 3). G The frequencies of M1 (CD11c⁺CD11b⁺F4/80⁺CD45⁺7AAD^-CD3^-CD19^-) and M2 (CD1206⁺CD11b⁺F4/80⁺CD45⁺7AAD^-CD3^-CD19^-) tumor-infiltrating macrophages were analyzed by flow cytometry. *p < 0.05. One-Way ANOVA test (n = 3-4). H The quantification of the M1/M2 ratio is shown. *p < 0.05. One-Way ANOVA test (n = 3). I A representative image of flow cytometric analysis of GzmB⁺ (GzmB^hiCD8⁺CD3⁺CD45⁺7AAD^-) T cells. J The frequency of GzmB⁺ (GzmB^hiCD8⁺CD3⁺CD45⁺7AAD^-) T cells was analyzed by flow cytometry. *p < 0.05. One-Way ANOVA test (n = 3-4). K The density of GzmB⁺ (GzmB^hiCD8⁺CD3⁺CD45⁺7AAD^-) T cells is shown. *p < 0.05. One-Way ANOVA test (n = 3-4). L The density of IFNγ⁺ (IFNγ^hiCD8⁺CD3⁺CD45⁺7AAD^-) T cells is shown (n = 3-4). *p < 0.05 and **p < 0.01. One-Way ANOVA test (n = 3-4). M The proposed mechanism of TGFβ1/TGFβR/Smad3/PRMT5-mediated ENO1 translocation for lactate release via MCT4.