Extended Data Fig. 7: tDC respond to microbial stimulation and uptake antigen in vivo.
a Splenic DC subsets purified as described in Figs. 1f and 4f were analyzed by Nanostring. Heatmap shows the expression of TLRs by each DC subset. N = 2 mice in 2 experiments. b Whole spleen cell suspensions were activated with LPS, MPLA, Resiquimod or PolyIC for 6 hrs. Heatmap showing the upregulation of MHCII and CD86 in pDC, tDC, ESAM+ EYFP+ and EYFP- DC2, and DC1. Min and max correspond to the lowest and highest gMFI per row, respectively. N = 6 mice per group. Data pooled from 4 experiments. c As in (b), but bar graphs (mean + SD) of cytokine production. Whole spleen cell suspensions were activated with LPS (n = 6), MPLA (n = 4), Resiquimod (n = 6) or PolyIC (n = 6) for 6 hrs. TNF and IL-12p40 were detected by intracellular cytokine staining. N represents mice and data was pooled from 4 experiments. Statistical differences were determined by One-way ANOVA with Tukey’s multiple comparisons test. d Mice were inoculated with yellow-green polystyrene (YG-PS) beads i.v. Three hrs later, spleens were harvested and the phagocytosis of the particle was assessed by flow cytometry in each DC population, gated as in Fig. 1f (n = 3 mice in 3 experiments). Bars represent mean + SD. Statistical differences were determined by One-way ANOVA with Tukey’s multiple comparisons test. e As in (d), but mice were inoculated with PKH26-labeled SRBC (n = 6 mice in 4 experiments).
