Fig. 8

In vivo immunogenic cell death induction and immunosuppressive TME reprogramming. Confocal microscopy images (A) and fluorescence intensity (B) of CRT protein exposure in tumor tissues treated with a) PBS; b) Fe-PHCN; c) Fe-PHCN@DOX; d) Fe-PHCN@DOX plus laser; and e) Fe-PHCN@DOX plus laser plus anti-PD-L1 nanobody. The quantification (C) and percentage (D) of matured DCs cells (CD11c⁺CD86⁺CD80⁺) by flow cytometric analyses after various treatments. Representative immunofluorescence images (E) and relative quantitative analysis (F) of hypoxia areas in tumors treated with different groups after staining with DAPI (blue) and anti-HIF-1α antibody (green). The quantification (G) and percentage (H) of M1-type macrophages (CD11b⁺F4/80⁺CD86⁺) by flow cytometric analyses after various treatments. The quantification (I) and percentage (J) of M2-type macrophages (CD11b⁺F4/80⁺CD206⁺) by flow cytometric analyses after various treatments. The quantification (K) and percentage (L) of CD8⁺ cytotoxic T (CD3⁺CD8⁺) cells by flow cytometric analyses after various treatments. M The percentage of CD4⁺ helper T (CD3⁺CD4⁺) cells by flow cytometric analyses after various treatments. N–P ELISA analysis of the levels of cytokines IL-6, TNF-⍺, and IFN-γ in serum of mice after various treatments. The scale bar represents 50 μm. The p values were analyzed using the Log-rank (Mantel-Cox) test. Data are presented as the mean ± standard error of the mean. *p < 0.05, **p < 0.01, ***p < 0.001