Extended Data Fig. 6 | Hemi-subdiaphragmatic vagotomy revealed
functional asymmetries of the vagus nerve. a–c, WT mice were subjected to
Sham, ventral (left) subdiaphragmatic vagotomy (LVx) or dorsal (right)
subdiaphragmatic vagotomy (RVx) (n = 4/group). Phenotypic analysis of T cell
and APC in colon was performed at day 2 after surgery. a, Frequency of Foxp3+
cells among CD4+ cells in colon. Representative contour plots. b, Frequency of
RORγt+ cells among Foxp3+ Treg in colonic LP. Representative contour plots
(left panel). Quantification (right panel). c, Frequency of ALDH+ cells among
MHC-II+ colonic APCs. Histograms of ALDH+ cells with colonic APCs (left panel).
Quantification (right panel). d–j, Ablation of sympathetic signalling via CG/
SMG does not affect maintenance of Treg in colon. d, Operative field for CG/
SMG ganglionectomy. e, Electrical activity in splanchnic nerve. The numbers in
parentheses correspond to the nerves indicated in d. f–i, WT mice were
subjected to Sham (n = 4) or ganglionectomy of CG/SMG (n = 5). Phenotypes of
immune cell in colon and spleen were analysed 2 days later. j, WT mice were
injected with MLA (α7-antagonist, 150 μg/day, i.p.) for 2 days and then splenic
T cells were analysed at 12 h after last injection (n = 5/group). f, i, j, Frequency of
Foxp3+ cells among CD4+ cells in colon (f) and spleen (i, j). g, Frequency of
RORγt+ cells among colonic Foxp3+ Tregs. h, Frequency of ALDH+ cells among
MHC-II+ colonic APCs. k–m, WT mice were subjected to Sham or HVx and then
injected daily with vehicle, salbutamol (β-agonist, 30 μg/day, i.p.) or
propanolol (β-antagonist, 300 μg/day, i.p.) for 2 days (n = 6/group). Frequency
of Foxp3+ cells among CD4+ cells (k), RORγ t+ cells among colonic Foxp3+ Tregs
(l), and ALDH+ cells among MHC-II+ colonic APCs (m) in the colon at 12 h after
the last injection. n, WT mice were subjected to Sham or HVx. T cell phenotypes
in colon, small intestine and spleen were analysed at day 2 after surgery.
Frequency of Foxp3+ cells among CD4+ T cells in the colon, the small intestine,
and the spleen (n = 4/group). Representative of two independent experiments
(a–n). P values obtained via one-way ANOVA with Tukey’s post hoc test (b, c, k–n)
and unpaired two-tailed Student’s t tests (f–j). Data are shown as mean ± s.e.m.