Figure 5. Transient Treg loss results in minimal skin inflammation.

Control wild-type (WT) or Foxp3^DTR mice were depilated and treated with DT according to the ‘early’ depletion protocol. (A) Total dermal infiltrate and (B) epidermal hyperplasia in DT treated control (WT + DT) and Tregs depleted (Foxp3^DTR + DT) dorsal skin on day 4 as measured by routine histology. (C) Representative H&E staining of skin from WT and Foxp3^DTR mice on day 4. (D) The absolute cell number of innate and adaptive immune cell subsets in skin as measured by flow cytometry. Single cell suspensions from day 1 skin were stimulated with PMA/ionomycin and the production of IL-22, IFNg, IL-17 and TNFalpha was assessed by flow cytometry. The proportion of cytokine producing (E) CD4⁺ Teff Cells, (F) CD8⁺ T cells, and (G) dermal γδ⁺ T cells in dorsal skin. (H) Flow cytometric quantification of Ki67⁺ bulge HFSCs at day 4 post-depilation in immune cell depleted and interferon-γ neutralized (or interferon-γ signaling deficient) mice. Scale Bars, 50 µm. Data are mean ± s.e.m. ns = no significant difference, Unpaired Student’s t-test (A, B); Two-way ANOVA (D–G); One-way ANOVA (H). Combined data from two experiments, with n = 3–4 mice per group. See also Figure S5.