Fig. 4: BSA-Man@Mn²⁺-Ft@Lap nanoassembly stimulates cGAS-STING-mediated dsDNA sensing machineries in DCs.

a Schematical illustration of the nanoassembly-stimulated cGAS/STING signaling of DCs in the TME. b Western blot analysis of the expression levels of STING, p-STING, TBK1, p-TBK1, IRF-3, and p-IRF-3 in BMDCs after the conditioning with supernatants from 4T1 cells treated with (I) control, (II) BSA-Man-Ft, (III) BSA-Man-Ft@Lap, (IV) BSA-Man@Mn²⁺-Ft, and (V) BSA-Man@Mn²⁺-Ft@Lap, respectively. c, d Flow cytometric analysis of dsDNA uptake and p-TBK1 levels in BMDCs incubated with 4T1 cells after different treatments. e Western blot analysis of the expression level of STING, p-STING, IRF-3, p-IRF-3, TBK1, and p-TBK1 in BMDCs after treatment with supernatants from 4T1 cells treated with BSA-Man@Mn²⁺-Ft@Lap, BSA-Man@Mn²⁺-Ft@Lap+DNase, and BSA-Man@Mn²⁺-Ft@Lap+HMGB1 antibody, respectively. f, g Flow cytometric analysis of dsDNA uptake and p-TBK1 levels in splenic cells co-cultured with 4T1 cells after different treatments. h cGAMP production in BMDCs after treatment with (I) control, (II) BSA-Man-Ft, (III) BSA-Man-Ft@Lap, (IV) BSA-Man@Mn²⁺-Ft, (V) BSA-Man@Mn²⁺-Ft@Lap, (VI) BSA-Man@Mn²⁺-Ft@Lap+DNase, and (VI) BSA-Man@Mn²⁺-Ft@Lap+HMGB1 antibody, measured by ELISA. Flow cytometry and western blot experiments in panels b–g were repeated three times independently with similar results. Data are presented as mean values ± SEM (n = 5 biologically independent samples for panel h). Statistical analysis was carried out via two-way ANOVA method. * Indicates significance at p < 0.05, *** indicates significance at p < 0.001, **** indicates significance at p < 0.0001. Source data are provided as a Source data file.