Extended Data Fig. 3: Peripheral Skint1 sustains steady-state DETC signatures of immunosurveillance.
From: Normality sensing licenses local T cells for innate-like tissue surveillance
a) Expression of Areg in DETCs from the combined dataset of single cell RNA sequencing of DETCs from wild-type FVB mice treated 7d prior with Skint1 Ab or isotype control by intradermal injection (n=1/group). b) Expression of selected Cluster 0 marker genes from the combined dataset as in a. c) Selected enriched gene ontology terms in each DETC cluster from the combined dataset as in a. d) Seurat cell cycle analysis of DETCs from the combined dataset as in a. e) Gene set enrichment analysis (GSEA) comparing Cluster 3 with other clusters in the combined dataset as in a, using publicly available gene set “GO positive regulation of immune response”. f) qPCR for cluster 3 marker gene expression within sorted 4-1BBhi and 4-1BBlo DETCs from untreated wild-type FVB mice (n=9). g) qPCR validation of differentially expressed genes in total DETC sorted from mice treated as in a (isotype n=9, Skint1 Ab n=8). h) Differentially expressed genes in Clusters 0 and 1 comparing Skint1 Ab and isotype treatments as in a. i) Expression of Ptpn6 in DETCs comparing Skint1 Ab and isotype treatments as in a. j) Flow cytometry of CD122 and CD45RB expression by DETCs from mice treated as in a (n=5/group). k) Flow cytometry of CD45RB expression by TCRγδ+ DETCs from wild-type FVB and FVB.Tac mice treated as in a (n=4/group). l) Flow cytometry of 4-1BBhi DETCs and CD122 expression by DETCs from wild-type FVB (n=7) and Langerin-DTA mice (n=5). Pooled data from two experiments (f-g) or representative of at least two experiments (j, l). Hypergeometric test with Benjamini-Hochberg correction (c), paired two-tailed t-test (f), unpaired two-tailed t-test (g, j, l), Wilcoxon test with Bonferroni’s correction (h), ordinary two-way ANOVA with Sidak’s multiple comparisons test (k). Error bars represent mean ± SD.
