Extended Data Fig. 5 | Afferent vagal, but not spinal cord, activation from
the liver is involved in colonic Treg homeostasis. a-e, WT mice underwent
either corn oil (Oil) or capsaicin (Cap) application to the hepatic vagal
branch. Phenotypic analysis of T cell and APC in colon was performed at
day 2 after application of capsaicin (c, d, n = 16/group; e, f, n = 8/group).
a, b, Representative f luorescence images of TRPV1 (red) and DAPI (blue) in NG
(a) and Th4-DRG (b). Scale bar indicates 100μm. c, Frequency of Foxp3+ cells
among CD4+ cells in colon. Representative contour plots (left panel).
Quantification (right panel). d, Frequency of RORγt+ cells among colonic
Foxp3+ Tregs. Representative contour plots (left panel). Quantification (right
panel). e, Frequency of ALDH+ cells among MHC-II+ colonic APCs. Histograms
of ALDH+ cells with colonic APCs (left panel). Quantification (right panel).
f, Expression of Aldh1a1 and Aldh1a 2 mRNA in colonic APC. g–p, Eight-week-
old WT type were intrathecally injected with capsaicin (g–j, n = 5/group)
and resiniferatoxin (RTX) (k–p, n = 4/group). TR PV 1+ nerves in spinal cord
(Th4-7 and Th13) and colonic immune cells were analysed at day 7 after
administration. g, Fluorescent immunohistochemistry of TRPV1+ neuron in
spinal cord (bar: 200μm). Frequency of Foxp3+ cells among CD4+ cells (h, n)
and RORγt+ cells among colonic Foxp3+ Tregs (i, o) in the colon. Frequency of
ALDH+ cells among MHC-II+ colonic APCs (j, p). k, l, Effects of intrathecal
injection of RTX in DRG (k) and NG (l). Scale bar indicates 100 μm. m, Colonic
CGRP levels. Representative of two independent experiments (a–b, g–o) or
pooled from two (e) or three (c, d, f) independent experiments. P values
obtained via unpaired two-tailed Student’s t tests. Data are shown as
mean ± s.e.m.