Journal: bioRxiv
Article Title: Colitis-primed circulating T cells drive small intestinal barrier function through Nod2, microbiota and myosin light chain kinase-dependent mechanisms
doi: 10.1101/2025.07.10.663215
Figure Lengend Snippet: (A) Percentages of CD4⁺ T cell subsets expressing RORγt and Gata3 in Foxp3⁻ or Foxp3 + CD4⁺ T cells isolated from the small intestine following colitis induction with TNBS, DSS, or oxazolone. (B) Representative flow cytometry plots of CD4⁺ T cells expressing RORγt, Foxp3, and Gata3 isolated from the small intestine of DSS-treated colitic mice and vehicle-treated controls. (C) Frequency of RORγt⁺Foxp3⁻CD4⁺ T cells in the small intestine of control and antibiotics (ABX)-treated WT mice following induction of colitis with TNBS, DSS, or oxazolone. (D) Duodenal and ileal paracellular permeability assessed by Ussing chamber in WT mice following CD4⁺ T cell depletion by monoclonal antibody or isotype control after induction of TNBS colitis. (E-J) Ileal paracellular permeability in: CD4 DTR mice treated with diphtheria toxin (DT) to deplete CD4⁺ T cells following TNBS (E) or oxazolone (F) colitis induction versus WT control mice treated with DT or PBS vehicle; (G) WT mice treated with FTY720 (a CD4⁺ T cell recirculation inhibitor) or vehicle; (H) RORγt −/- mice and WT littermate controls following TNBS or oxazolone-induced colitis; (I) CD4 Cre RORγt flox mice and RORγt flox littermate controls; and (J) Foxp3 Cre RORγt flox mice and RORγt flox littermate controls, all following TNBS or oxazolone-induced colitis. Each data point represents one mouse; n ≥ 3 per group. Data are presented as mean ± s.e.m. from at least three independent experiments. All p- values were calculated using one-way ANOVA. Statistical significance is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. indicated group; ns, not significant.
Article Snippet: Intracellular staining was performed with Alexa Fluor (AF)700-conjugated Foxp3 (clone FJK-16s, ThermoFisher) or with AF488-conjugated Gata3 (clone TWAJ, ThermoFisher) and AF647-conjugated RORγt (clone Q31-378, BD).
Techniques: Expressing, Isolation, Flow Cytometry, Control, Permeability
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