Supplementary Figure 7: Depletion of T_RM cells via NAD.

(a) Flow cytometry of P2X7-Art2.2-coexpression. Representative control stainings for Fig. 7f showing the expression of P2X7 and Art2.2 in spleen CD3^- and CD3⁺ cells. The frequency of double-positive cells is indicated. (b) Frequency of CD44⁺CD69^- effector memory T cells (T_EM) in mice treated with or without NAD. n = 22. (c) Representative (left) and quantitative (right) flow cytometry of CD1d tetramer⁺ NKT cells in the colon of mice treated with or without NAD as well as representative gating control flow cytometry of CD1d tetramer⁺ NKT cells in the liver (far right). n = 14. (d) Frequency of CD3⁺ T and CD1d tetramer⁺ NKT cells among lymphoid LPMCs after treatment with PBS. n = 8. (e) Immunohistochemistry staining for CD4 (n = 16) and MPO (n = 17) in colon tissue of WT mice with TNBS colitis after treatment with or without NAD from three independent experiments. (f) Flow cytometry of Art2.2 expression in T cells versus non-T cells. Left: Representative histograms of Art2.2 expression on CD4⁺ T cells, CD3^- non T cells and fluorescence minus one (FMO) control for Art2.2. Right: Quantitative expression. n = 10 mice from three independent experiments. (b-f) Boxplots – median with upper and lower quartile, whiskers – min and max. n.s. – not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, paired (d, f) or unpaired (b, c, e) two-tailed student’s t-test.