Fig. 5.

The up-regulation of CD103 on splenic cDC1s in response to activation of NOD2 is dependent on GM-CSF. (A) Flow cytometry analysis of CD103 expression on splenic CD8α⁺ cDC1s collected from C57BL/6 littermates 16 h postinjection with GM-CSF (200 ng i.p.). (B) Flow cytometry analysis of CD103 expression on splenic CD8α⁺ cDC1s collected from GM-CSF–deficient mice (Csf2^−/−) and their heterozygote littermates (Csf2^+/−) 16 h postinjection with PBS or MDP (50 μg i.p.). (C) qPCR analysis of Csf2 gene expression in the parietal peritoneum and spleen of C57BL/6 mice 4 or 24 h post-MDP treatment (50 μg). (D) CD90.1⁺ OT1 cells were adoptively transferred into CD90.2⁺Csf2^−/− mice and their CD90.2Csf2^+/− littermate controls and treated as in the scheme outlined in Fig. 1D. The proportion of donor-derived CD8 T cells (D), as well as the proportion of donor-derived T cells expressing markers of short-lived effector cells (E; CD127⁻KLRG1⁺) was determined by flow cytometry. Individual mice from one representative experiment (A; C, Right; and D) or two pooled litters (B and C, Left) are shown. Horizontal bars denote the mean, and error bars denote SEM. Asterisks denote significantly different from PBS control as determined by Student’s t test (A), genotype-matched PBS-treated littermate control as determined by two-way ANOVA followed by Tukey’s multiple comparisons test (B), or naïve mice as determined by one-way ANOVA followed by Tukey’s multiple comparisons test (C). Asterisks and horizontal brackets denote significant genotype-dependent differences regardless of treatment as calculated by two-way ANOVA; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001; NS, no significant difference between linked groups.