Fig. 3: Effective DC maturation in vitro by SCCNVs.

a In vitro uptake of CCNVs and SCCNVs by DCs for 10 min, as evaluated by flow cytometric analysis. n = 3. b mRNA levels of DC maturation markers (IL-6 and IL-12p40) in BMDCs treated with PBS, CCNV, SCCNV, or LPS for 4 h in vitro, as evaluated by qRT‒PCR. n = 3-4. c Representative flow cytometry plot and percentage of CD80 and CD86 double-positive cells in BMDCs treated with PBS, CCNV, SCCNV, or LPS for 24 h in vitro, evaluated by flow cytometric analysis. n = 4. d Flow cytometric analysis of MHC class I expression of BMDCs treated with PBS, CCNVs, SCCNVs, or LPS for 24 h in vitro, as evaluated by flow cytometric analysis. n = 3. e Flow cytometric analysis showing that the enhanced DC maturation induced by SCCNVs is likely due to IFN-γ and TNF-α contained in SCCNVs. Exogenous IFN-γ and TNF-α were added to the CCNV + IFN-γ + TNF-α group. n = 4. f Effects of various treatments for 24 h on DC maturation, as evaluated by flow cytometric analysis. siRNA-SCCNVs are nanovesicles derived from senescent cancer cells transfected with siRNA specific for IFN-γ and TNF-α. n = 4. In (b–f), LPS was used as the positive control. Data represent the mean ± SD. Statistical significance was calculated by Student’s t-tests or one-way ANOVA with Tukey’s multiple comparisons test. *P < 0.05 versus PBS; †P < 0.05 versus CCNVs; ‡P < 0.05 versus SCCNVs in b, c, d and f and versus CCNVs + IFN-γ, TNF-α in e. ns not significant.