Figure 6.

STING Activation Induced DC Maturation and Promoted the Interaction between LYN and STING in DC

(A–D) Fluorescent western blot shows (A) the immunoprecipitation (IP) with STING-N (red) and blots with Lyn (green) and (B) cell lysate of activated BMDC with DMXAA at 0 and 3 h (C) A reverse IP using the Lyn antibody and blot with STING antibody and (D) cell lysate of activated BMDC with DMXAA at 0 and 3 h. Data show a representative of four experiments.

(E–L) (E) Western blot analysis of Sting-activated BMDC with or without PP2 inhibitor showed the phosphorylation of Lyn (Try507) and Akt (Ser473). Data are representative of three mice per group. Sting-activated BMDCs were cultured with Lyn inhibitor (PP2) and analyzed by (F–H) flow cytometry shows the percentage of (F) CD80⁺CD11c⁺, (G) I-Ab⁺CD11c⁺, and (H) PDCA⁺CD11c⁺ cells (N = 3 per group), and (I–L) the relative RNA expression (normalized by actin) of (I) Irf3, (J) Irf7, (K) Isg15, and (L) Cxcl10 are shown (N = 4 per group).

(M and N) Confocal microscopy of DMXAA-activated BMDC from WT, Sting^gt/gt, and Fcgr2b^−/−. Sting^{wt/gt -} mice for 6 h. (M) The quantification of colocalization signals between STING and Lyn (N = 5 per group). Data are shown as mean ± SEM; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001. (N) Immunofluorescence staining of BMDC shows Lyn (green), STING (red), and DAPI (blue) (scale bar, 20 μm). Data show a representative of five experiments.