Extended Data 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) qPCR expression of Skint1 and Skint2 in KCs sorted from wild-type C57BL/6 mice at indicated timepoints following UVR (n=3/timepoint). b) Flow cytometry frequency of activated LCs following UVR of ear skin in wild-type C57BL/6 mice (n=3/timepoint). c) Flow cytometry of CD69 expression by DETCs following UVR of ear skin in wild-type C57BL/6 mice (n=3/timepoint). d) Flow cytometry analysis of fold change in CD69 gMFI on DETCs 24hr post-UVR relative to unchallenged skin from wild-type FVB (n=25), FVB.Tac (n=13), Trgv5-/-Trdv4-/- (n=13) and Skint1-transgenic FVB.Tac mice (n=6). e) qPCR for effector genes in DETCs sorted from wild-type C57BL/6 mice at 24hr post-UVR, following pre-treatment with intradermal injection of Skint1 Ab or isotype control (isotype normal n=8, Skint1 Ab normal n=8, isotype UVR n=10, Skint1 Ab UVR n=10). f) Flow cytometry frequency of activated LCs from mice treated as in e) (n=7/group). Flow cytometry of GFP expression by DETCs from ear sheets of Nur77-GFP mice which were cultured for 4hr in medium containing g) anti-Skint1 or isotype control antibodies and h) PP2 or DMSO vehicle prior to UVR, and analyzed 24hr later (n=3/group). Pooled data from 4 (d) or 2 (e) independent experiments, or representative of 3 experiments (g-h). Ordinary one-way ANOVA with Dunnett’s multiple comparisons test relative to 0hr (a) or wild-type (d), paired two-way ANOVA with Sidak’s multiple comparisons test (e, f-h). Error bars represent mean ± SD.
