Fig. 4: DETC sensing of genotoxic epithelial stress requires acute exposure to Skint1.
From: Normality sensing licenses local T cells for innate-like tissue surveillance
a, Confocal imaging of CD3+ DETCs (cyan), F-actin within a CD3+ mask (yellow) and γH2A.X+ nuclei (magenta) following UVR of ear skin of wild-type C57BL/6 mice. Data are representative of n = 5 per time point; scale bar, 20 μm. b, Scaled gene expression from RNA sequencing of DETCs isolated from ear skin 24 h after UVR of wild-type C57BL/6 mice (n = 3 per group); VST, variance-stabilizing transformation. c, GSEA of UVR-induced gene expression changes in DETCs using signatures of steady-state DETC clusters from Fig. 3. d, qPCR for DETC gene expression in the epidermis of FVB wild-type (n = 15), Tac (n = 9), Trgv5–/–Trdv4–/– (n = 5) and Skint1-transgenic FVB.Tac (n = 9) mice 24 h after UVR. e, qPCR for effector genes in DETCs sorted at day 1 after UVR from wild-type C57BL/6 mice pretreated with i.d. injection of Skint1 antibody or isotype control (isotype control normal, n = 8; Skint1 antibody normal, n = 9; isotype control/Skint1 antibody UVR, n = 10 per group). f, Flow cytometry of 4-1BB and CD122 expression in DETCs from wild-type C57BL/6 mice treated as in e (n = 8 isotype control; n = 9 Skint1 antibody). g, Flow cytometry of GFP expression in DETCs following UVR of Nur77–GFP reporter mice (n = 3 per time point). h, Flow cytometry of GFP expression in DETCs from Nur77–GFP reporter mice treated as in e (n = 7 per group). Data are pooled data from two (e and f) or three (d) independent experiments or are representative of two (a) or three (h) independent experiments. A paired (d, f and h) or ordinary (e) two-way ANOVA with Sidak’s multiple comparisons test was performed. Error bars represent mean ± s.d.
