Fig. 3. Tonic TNFR2 signaling is required for the presence of the R2D2 population in the lung.

a–c Flow cytometry analysis of TNFR2 a and pRelB expression b on lung cDC2 in TNFR2^fl/fl and TNFR2^fl/flCD11c^cre mice at steady state. WT mice were treated i.n. with anti-TNFR2 blocking antibody (20 μg) or TNFR2-Fc recombinant protein (2 μg). Lungs were harvested 24 h later; n = 3–4 mice/group. Data are representative of two independent experiments. d Quantification of Mgl2⁺TNFR2⁺ cDC2 in the lungs of indicated groups; n = 3–4 mice/group. Data are representative of two independent experiments. e, f Flow cytometry analysis of TNF expression by anti-TNF mAb (clone D2D4) e or mouse TNFR2-Fc recombinant protein f; n = 3 mice/group. Data are representative of three independent experiments. g–h Flow cytometry analysis of TNFR2⁺ cDC2 g and Ki67 expression h in mice treated i.n. with anti-TNFR2 mAb (TR75.89) and TNFR2-agonist TNF_D221N/A223R (1 μg). Lungs were harvested 24 h later; n = 3 mice/group. Data are representative of two independent experiments. Graphs represent the mean with error bars indication s.e.m. P values determined by one-way ANOVA Tukey’s multiple comparison test c, d or unpaired student t-test g, h. *P < 0.05, **P < 0.001, and ***P < 0.0001.