Fig. 3: BSA-Man@Mn²⁺-Ft@Lap nanoassembly induced immunogenic apoptosis of tumor cells via Lap-mediated chemotherapy.

a, b Fluorescence imaging on the endocytosis of BSA-Man@Mn²⁺-Ft-FITC@Lap by 4T1 tumor cells after incubation under different pH values for 12 h and 24 h. c Quantitative analysis of FITC fluorescence from panel a/b. d Flow cytometric analysis regarding the accumulation of mannose-modified BSA contents in DCs after 24 h of incubation with (I) control, (II) BSA-FITC@Mn²⁺-Ft@Lap at pH 7.4, (III) BSA-FITC-Man@Mn²⁺-Ft@Lap at pH 7.4, (IV) BSA-FITC@Mn²⁺-Ft@Lap at pH 6.5, and (V) BSA-FITC-Man@Mn²⁺-Ft@Lap at pH 6.5. e CLSM imaging of HMGB1 immunofluorescence in 4T1 cells after treatment with (I) control, (II) BSA-Man-Ft, (III) BSA-Man-Ft@Lap, (IV) BSA-Man@Mn²⁺-Ft, and (V) BSA-Man@Mn²⁺-Ft@Lap. Greater red fluorescence indicates lower HMGB1 release. f Quantitative fluorescence analysis of HMGB1 release in panel (e). g Western blot analysis on the expression levels of HMGB1, CRT and apoptosis markers (Bax, Bcl-2) in 4T1 cells with different treatments. h CLSM imaging of CRT expression after different treatments with (I) control, (II) BSA-Man-Ft, (III) BSA-Man-Ft@Lap, (IV) BSA-Man@Mn²⁺-Ft, and (V) BSA-Man@Mn²⁺-Ft@Lap. Greater red fluorescence indicates higher expression levels. i Quantitative fluorescence analysis of CRT in panel (h). j ATP levels in cell culture supernatants after 24 h of different treatments: (I) control, (II) BSA-Man-Ft, (III) BSA-Man-Ft@Lap, (IV) BSA-Man@Mn²⁺-Ft, and (V) BSA-Man@Mn²⁺-Ft@Lap. Data are presented as mean values ± SEM (n = 3 biologically independent samples for panels c, f, i, and j). Flow cytometry and western blot experiments in panels d and g were repeated three times independently with similar results. Statistical analysis for panels c, f, i, and j were carried out via multiple comparisons one-way ANOVA method. * Indicates significance at p < 0.05, ** indicates significance at p < 0.01, *** indicates significance at p < 0.001, **** indicates significance at p < 0.0001. Source data are provided as a Source data file.