Extended Data Fig. 9 | The effects of gut-microbiota on colonic Treg
maintenance of the liver-brain-gut axis. a–c, WT mice were subjected to
Sham or HVx (n = 4/group). Faecal samples were collected on pre-treatment
and days 2 post-surgery from the identical mice. a, α-diversity in the faecal
microbiota. b, Principal coordinate analysis (PCoA) based on the weighted
UniFrac analysis of bacterial community structures (black, pre-treatment; red,
sham; blue, HVx). The two components of the weighted PCoA plot explained
45% and 22% of the variance. Dissimilarities between two groups were
evaluated by permutational multivariate analysis of variance (PERMANOVA).
c, Phylum-level taxonomic distribution. d, e, Faecal samples derived from
sham-operated or HVx mice were inoculated into five-week-old male GF mice,
followed by immunological phenotyping at day 21 post-inoculation (n = 5/
group). f, g, WT mice were subjected to Sham or HVx and co-housed in our SPF
facility for 2 days. Graphs show pooled data of three independent experiments
(n = 14/group). h–k, At day 14 post-parabiosis, mice were subjected to sham or
HVx (n = 10/group). Phenotypes of colonic immune cell were analysed 2 days
later (h). l, m, WT mice were treated with Abx-cocktail (metronidazole,
vancomycin, ampicillin and neomycin) for 3 weeks and subjected to Sham or
HVx. Colonic T cell phenotypes were analysed 2 days later. Graphs show pooled
data of three independent experiments (n = 10/group). d, f, i, l, Frequency of
Foxp3+ cells among CD4+ T cells in colon. e, g, j, m, Expression of RORγt+ in
colonic Foxp3+ Tr e g. k, Frequency of ALDH+ cells among MHC-II+ colonic APCs.
P values obtained via one-way ANOVA with Tukey’s post hoc test (a, d, e, l, m) or
unpaired two-tailed Student’s t tests (f, g, i–k). Data are shown as mean ± s.e.m.