Fig 1: S1PR2 blockade in NHEK increases psoriasis-related inflammation. (A) Representative immunofluorescence images of TLR7 (green) and nuclei (blue) of NHEKs. Left panel shows NHEKs stained with DAPI and secondary antibody. Right panel shows NHEKs stained with DAPI, primary, and secondary antibody (B) mRNA expressions of TNF-α, Il-17C, IL-1β, and (C) mRNA expressions of MYD88, TRAF6, IRF7, NFKB1, and RELA of NHEKs treated with PBS for 24 hours or with IMQ for 2, 5, and 24 hours. Before IMQ treatment, S1PR2 was inhibited by incubating NHEKs with 10 μM JTE013 for 2 hours. Each gene expression was measured with RT-qPCR and normalized to GAPDH mRNA expression level. Data shown are the mean ± SD (n = 3) and are representative of three independent experiments with similar results. ****p < 0.0001, ***p <0.0005, **p < 0.005. ns, not significant.
Fig 2: S1PR2 deletion increases MyD88/NF-κB pathway molecules but not affects TLR7 expression. mRNA expressions of Tlr7, Myd88, Traf6, Irf7, Nfkb1, and Rela of (A) Balb/c and S1pr2-/- mouse epidermis, and (B) S1pr2fl/fl and S1pr2fl/fl K14-Cre mouse epidermis were measured with RT-qPCR and normalized to Gapdh mRNA expression level. (C) The protein level of NF-κB was measured by enzyme-linked immunosorbent assay. Data shown are the mean ± SD (n = 3) and are representative of three independent experiments with similar results. ****p < 0.0001, ***p <0.0005, **p < 0.005, *p < 0.05. ns, not significant.
Fig 3: Spatial transcriptomics reveals that S1PR2 deletion in keratinocytes results in psoriasis-like inflammation in the epidermis. (A) Graph-based clustering (upper lane) and UMAPs (lower lane) of S1pr2 fl/fl and S1pr2 fl/fl K14-Cre mouse epidermis after IMQ administration. (B) Spatial gene expression analysis of Tnf, Il-17a, Il-17c, Il-17f, Il-12a, Il-12b, Il-23a, Il-27, Tlr7, Myd88, Nfkb1, Rela, and Il-1β in S1pr2 fl/fl and S1pr2 fl/fl K14-Cre mouse epidermis after IMQ administration. (C) Representatives of pathway enrichment analysis of the mRNA levels by spatial transcriptomics of S1pr2 fl/fl and S1pr2 fl/fl K14-Cre mouse epidermis after IMQ administration. Genes upregulated and downregulated in S1pr2 fl/fl K14-Cre mouse epidermis are highlighted in green and red, respectively. Fold change > 2 and adjusted P value < 0.05.
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