Fig. 2: Ag+ DC express high levels of activation markers and transcripts associated with Th cell polarization.

C57BL/6 mice were immunized i.d. with fluorescently labeled Ms, Nb, Ca, or PBS as a control. Flow cytometry (a–d) and cell sorting (e) were performed on LN cell suspensions 2 days later. Bar graphs show mean median fluorescence intensity (MFI) ± SD, each symbol corresponds to one mouse. a–d Expression of CD40, CD86, PDL2, and CCR7 across Ag+ and Ag− DC subsets from  PBS-injected or immunized mice. Group sizes are; n = 3 (CD40 PBS), 4 (CD86 immunized, PDL2 Ms and PDL2 Nb) or 5 (CD40 immunized, CD86 PBS, PDL2 PBS, PDL2 Ca and all CCR7) female mice from 1 of 2 (CD40, PDL2, and CCR7) or 3 (CD86) independent experiments that gave similar results. Statistical significance was assessed using Two-Way ANOVA with Sidak’s post-test. Hash symbols above individual columns refer to comparisons to the PBS group. NS: not significant; **^,##p < 0.01; ***^,###p < 0.001; ****^,####p < 0.0001. Exact values are shown for 0.05 > p > 0.01. e Dynabead-enriched DC were sorted into Ag− and Ag+ subsets for RT-qPCR analysis. Heatmaps show differential expression analysis (log2 fold change) of transcripts in DC subsets from immunized mice compared to PBS controls. Data refer to three biological replicates each containing cells sorted from pooled LN from five female mice. Statistical significance was assessed using One-Way ANOVA with Holm–Sidak’s post-test; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source Data file.