Fig. 2: Activation of dendritic cells induced by YCW NPs and its mechanism.

A–B Activation of BMDCs after BMDCs incubation with different sizes YCW NPs (including large NPs, middle NPs, and small NPs) and LPS (positive control) for 24 h. A Representative dot plots of co-stimulatory molecules CD80 and CD86 expression on BMDCs and (B) corresponding quantification of BMDCs maturation (n = 3). C–F Concentration of pro-inflammatory cytokines secreted by BMDCs after incubation with different sizes YCW NPs as indicated (n = 3). C TNF-α; D IL-12p70; E IL-1β; F IL-6. G–L Representative Western blotting result of Dectin-1/Syk pathway and TLR2/MyD88 pathway from proteins of BMDCs after incubation with three YCW NPs and LPS for 24 h (n = 3), including (H) TLR2, (I) p-Syk, (J) p-P65, (K) MyD88, (L) Dectin-1. M–P After utilizing Dectin-1 competitor laminarin for 2 h, representative western blotting result of Dectin-1/Syk pathway from proteins of BMDCs after incubation with small NPs for 24 h (n = 3), including (N) p-Syk, (O) p-P65, (P) Dectin-1. Q–T After utilizing TLR2 inhibitor C29 for 2 h, representative western blotting result of TLR2/MyD88 pathway from proteins of BMDCs after incubation with small NPs for 24 h (n = 3), including (R) p-P65, (S) TLR2, (T) MyD88. U A scheme revealing the mechanism of YCW NPs to activate dendritic cells via a Dectin-1 and TLR2-mediated manner. All experiments were run in triplicate. Statistical significance between different groups was obtained by Student’s t tests (two-tailed) (N–P, R–T) and one-way ANOVA using the Tukey post-test (B, C–F, H–L). ****P < 0.0001; ***P < 0.005; **P < 0.01; *P < 0.05. Data are means ± SD. BMDCs: bone marrow-derived dendritic cells, LPS: lipopolysaccharides. The samples for western blotting analysis derived from the same experiments and the blots were processed in parallel.