Fig. 3: Uptake of MiBaMc and ICD in tumor cells triggered by MiBaMc.

a CLSM images showing the intracellular uptake of free Ce6, BaMc, and MiBaMc in 4T1 cancer cells after 8 h incubation. Blue, nucleus stained with Hoechst 33342; red, Ce6 fluorescence; green, mitochondria stained with Mito-Tracker green. Scale bar, 25 μm. The experiments in a were repeated three times with similar results. b Relative viability of 4T1 cancer cells after treatment with different formulations (n = 4 independent experiments). c Flow cytometry analysis of the mitochondrial membrane potential of 4T1 cells after treatment with different formulations. d Corresponding green fluorescence intensity of different groups based on the flow cytometry results in c (n = 3 independent experiments). Statistical analysis was conducted by one-way ANOVA with Tukey’s tests. ^*P < 0.05, ^***P < 0.001^{, ****}P < 0.0001. CLSM images showing e CRT exposure and f HMGB1 release in 4T1 cells after each treatment. Blue, nucleus stained with Hoechst 33342; Green, CRT stained with mouse anti-CRT primary antibody and then with Alexa Flour-488 conjugated goat anti-mouse secondary antibody; Red, HMGB1 stained with rabbit anti-HMGB1 primary antibody and then with Alexa Flour-647 conjugated goat anti-rabbit secondary antibody. Scale bar, 25 μm. g Western blot assay of HMBG1 and CRT proteins expressed in 4T1 cells after treatment with each formulation. The experiments in e-g were repeated three times with similar results. Quantification of extracellular release of h HMGB1 and i ATP in culture medium after different treatments (n = 3 independent experiments). Statistical analysis was conducted with ANOVA with Tukey’s tests. ^*P < 0.05, ^**P < 0.01. ^***P < 0.001, ^****P < 0.0001. j Flow cytometry analysis of the matured DCs (CD80⁺CD86⁺) after different treatments (n = 3). k Quantification of the percentage of matured DCs after different treatments (n = 3 independent experiments). Statistical analysis was conducted with ANOVA with Tukey’s tests. ^**P < 0.01. ^***P < 0.001, ^****P < 0.0001. Data are presented as mean values ± SD. Source data are provided as a Source Data file.