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nature research | reporting summary
October 2018washed twice in PBS before digestion with 1.0 mg/ml collagenase (Wako) and 0.05 mg/ml DNase (Sigma-Aldrich) for 60 min at
37°C. LP suspensions were passed through a 70-ђm filter. Cells were then separated with a Percoll density gradient (GE
Healthcare, IL, USA). The numbers of live cells were determined by Countess II (Thermo Fisher Scientific).Instrument Isolated cells were stained with fluorochrome-conjugated antibody.
For cell sorting: FACSAria II (Becton Dickinson).
For analysis: FACSCanto II (Becton Dickinson).
Software FACSDiva for collection and FlowJo (v10) for analysis
Cell population abundance Populations were validated for purity by a post-sort analysis by FACS
Gating strategy Gating strategies are reported in method section and Extended Data 3b.
Colonic and splenic APCs in mice: 7AAD-CD45.2+CD3-B220-NK1.1-MHC-II+
Colonic CD11c single postive cells (CD11c-SP): 7AAD-CD45.2+CD3-B220-NK1.1-MHC-II+CD11b-CD11c+
Colonic CD11c CD11b double postive cells (DP): 7AAD-CD45.2+CD3-B220-NK1.1-MHC-II+CD11b+CD11c+
Colonic CD11b single postive cells (CD11b-SP): 7AAD-CD45.2+CD3-B220-NK1.1-MHC-II+CD11b+CD11c-
Colonic total Treg in mice: FVD-CD45.2+CD3+CD4+Foxp3+
Colonic peripheral Treg: FVD-CD45.2+CD3+CD4+Foxp3+RORgt+Helios-
Colonic Th1: FVD-CD45.2+CD3+CD4+IFNg+
Colonic Th2: FVD-CD45.2+CD3+CD4+Gata3+
Colonic Th17: FVD-CD45.2+CD3+CD4+RORgt+IL17A+
Colonic APCs in human: CD45+CD3-CD19-CD56-HLA-DRhi
Neuro-Spheroid derived neuron: GFP+CD45.2-CD184-CD44-CD24+
Colonic total Treg in rat: FVD-CD3+CD4+Foxp3+
Tick this box to confirm that a figure exemplifying the gating strategy is provided in the Supplementary Information.