Figure S5.

IFN-γ expression and CD8 T cells drive LN expansion. (a and b) Efficacy of anti-Ly6G antibody-mediated neutrophil depletion (n = 3–6 mice from two independent experiments; a) or anti-NK1.1 antibody-mediated NK cell depletion (n = 5 mice from two independent experiments; b); unpaired Student’s t test. (c) Counts of lymphocytes and neutrophils in popLNs in diphtheria toxin–treated Clec9aCre:Rosa-iDTR or iDTR mice 2 d after unilateral sciatic denervation; n = 3–4 mice; data are representative of two independent experiments; two-way ANOVA; Šídák’s post-test. (d) Efficacy of diphtheria toxin–mediated Clec9aCre:Rosa-iDTR–specific cell depletion. Counts of total cells and DC subsets in spleen in diphtheria toxin–treated Clec9aCre:Rosa-iDTR or control mice 2 d after unilateral sciatic denervation; n = 3–4 mice; data are representative of two independent experiments; one-way ANOVA; Tukey’s post-test. (e) Counts of indicated subsets in popLNs 1 wk after unilateral sciatic denervation with or without IFN-γ blockade; n = 5 mice; data are representative of two independent experiments; two-way ANOVA; Šídák’s post-test. (f) Percentage of IFN-γ⁺ cells in indicated subsets; n = 5 mice; data are representative of two independent experiments; unpaired Student’s t test. (g) Counts of indicated subsets in popLNs 1 wk after unilateral sciatic denervation with or without CD8 depletion; n = 5 mice; data are representative of two independent experiments; two-way ANOVA; Šídák’s post-test. (h) Percentage of IFN-γ⁺ cells in indicated subsets from popLNs 1 wk after unilateral sciatic denervation with PBS or clenbuterol treatment; n = 5 mice; data are representative of two independent experiments; two-way ANOVA; Šídák’s post-test. (i) Presence of sympathetic TH⁺ nerves close to CD8 T cells in the popLN. B cell follicles are highlighted with dotted lines. Scale bar, 50 µm. (j) Percentage of viable isolated CD8 T cells after in vitro restimulation with PMA/ionomycin (P/I) in the presence or absence of clenbuterol; n = 7 mice from two independent experiments. Clec9aCre:Rosa-iDTR and iDTR mice were 8–12 wk old and bred in the animal facility. The other mice in this figure were 8–12-wk-old WT males purchased from Charles River or Janvier Labs. All data are presented as mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Clen, clenbuterol; DT, diphtheria toxin; Mϕ, macrophage; Sx, sciatic denervation.