Optimal maturation of CD8α+αDCs after CD1d-mediated activation of iNKT cells combined with CpG-ODN. Mice transferred with Vα14-Jα18 and OT-I splenocytes were immunized i.m. with OVA peptide alone or in combination with CpG or CD1d-fus/CpG followed or not by systemic treatment with the CD1d-antitumor protein. Mice were bled after 6 hours and sacrificed 20 hours after i.m. immunization. A. Twenty hours post immunization, splenic DC subsets were discriminated according to the gating strategy described in Additional file 1: Figure S1, and each subset was analyzed for the expression of CD40 and CD86 maturation markers. Data are shown as histogram overlays of the different treatment groups as indicated. B. Mean fluorescence intensity of CD40 on gated CD8α+DCs. C. Mean fluorescence intensity of CD86 on gated CD8α+DCs. D. Mean fluorescence intensity of CD70 on gated CD8α+DCs. E. Sera from mice were collected 6 hours after the immunization and IL-12p70 was measured using a BD flex set assay. Bar graphs show mean of fluorescence or cytokine level in pg/ml as mean +/α SEM for 3 samples per group. Data are representative of 2 independent experiments. *, p <0.05; **, p <0.01; ***, p <0.001; ****, p <0.0001.