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EpSC proliferation (S-phase EdU labeling) following 24h pharmacological application of PG inhibitors [Indo (Indomethacin; COX non-selective inhibitor), iCOX-1 (SC-560; selective COX-1 inhibitor), iEP4 (ONO-AE3-208; EP4 receptor antagonist)] or PG enhancers [i15-PGDH <t>(SW033291;</t> <t>15-PGDH</t> inhibitor), dmPGE 2 ]. One-Way ANOVA with Šidák multiple-comparison correction. (C) Experimental schematic for genetic perturbation of LCs. (D-E) Representative IF image and COX-1 MFI of LCs from LV expressing gCtrl or gPtgs1. (F) EpSC proliferation (S-phase EdU labeling) and epidermal thickness in gCtrl or gPtgs1 animals. (G) Experimental schematic of in utero LV delivery for genetic perturbation of EpSCs. (H) Volcano plot of differential gene expression between gCtrl or gPtger4 EpSCs. Genes associated with proliferation, differentiation and barrier function highlighted. (I) Heatmap showing relative Z-score normalized expression of key genes related to epidermal stress and proliferation in gPtger4 or gCtrl EpSCs. (J) Pathway-level analysis using Reactome database in upregulated genes in gPtger4 EpSCs. (K) Representative IF images and quantification of EpSC proliferation (S-phase EdU labeling) in gCtrl or gPtger4 LV + regions. (L) Representative H&E staining and quantifications of epidermal thickness in gCtrl or gPtger4 (transduced with 70% efficiency). For (A-L): Each dot corresponds to data from one mouse with median shown, n ≥ 3 mice per condition. Unless otherwise stated: two-tailed unpaired Student’s t tests.
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EpSC proliferation (S-phase EdU labeling) following 24h pharmacological application of PG inhibitors [Indo (Indomethacin; COX non-selective inhibitor), iCOX-1 (SC-560; selective COX-1 inhibitor), iEP4 (ONO-AE3-208; EP4 receptor antagonist)] or PG enhancers [i15-PGDH <t>(SW033291;</t> <t>15-PGDH</t> inhibitor), dmPGE 2 ]. One-Way ANOVA with Šidák multiple-comparison correction. (C) Experimental schematic for genetic perturbation of LCs. (D-E) Representative IF image and COX-1 MFI of LCs from LV expressing gCtrl or gPtgs1. (F) EpSC proliferation (S-phase EdU labeling) and epidermal thickness in gCtrl or gPtgs1 animals. (G) Experimental schematic of in utero LV delivery for genetic perturbation of EpSCs. (H) Volcano plot of differential gene expression between gCtrl or gPtger4 EpSCs. Genes associated with proliferation, differentiation and barrier function highlighted. (I) Heatmap showing relative Z-score normalized expression of key genes related to epidermal stress and proliferation in gPtger4 or gCtrl EpSCs. (J) Pathway-level analysis using Reactome database in upregulated genes in gPtger4 EpSCs. (K) Representative IF images and quantification of EpSC proliferation (S-phase EdU labeling) in gCtrl or gPtger4 LV + regions. (L) Representative H&E staining and quantifications of epidermal thickness in gCtrl or gPtger4 (transduced with 70% efficiency). For (A-L): Each dot corresponds to data from one mouse with median shown, n ≥ 3 mice per condition. Unless otherwise stated: two-tailed unpaired Student’s t tests.
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EpSC proliferation (S-phase EdU labeling) following 24h pharmacological application of PG inhibitors [Indo (Indomethacin; COX non-selective inhibitor), iCOX-1 (SC-560; selective COX-1 inhibitor), iEP4 (ONO-AE3-208; EP4 receptor antagonist)] or PG enhancers [i15-PGDH <t>(SW033291;</t> <t>15-PGDH</t> inhibitor), dmPGE 2 ]. One-Way ANOVA with Šidák multiple-comparison correction. (C) Experimental schematic for genetic perturbation of LCs. (D-E) Representative IF image and COX-1 MFI of LCs from LV expressing gCtrl or gPtgs1. (F) EpSC proliferation (S-phase EdU labeling) and epidermal thickness in gCtrl or gPtgs1 animals. (G) Experimental schematic of in utero LV delivery for genetic perturbation of EpSCs. (H) Volcano plot of differential gene expression between gCtrl or gPtger4 EpSCs. Genes associated with proliferation, differentiation and barrier function highlighted. (I) Heatmap showing relative Z-score normalized expression of key genes related to epidermal stress and proliferation in gPtger4 or gCtrl EpSCs. (J) Pathway-level analysis using Reactome database in upregulated genes in gPtger4 EpSCs. (K) Representative IF images and quantification of EpSC proliferation (S-phase EdU labeling) in gCtrl or gPtger4 LV + regions. (L) Representative H&E staining and quantifications of epidermal thickness in gCtrl or gPtger4 (transduced with 70% efficiency). For (A-L): Each dot corresponds to data from one mouse with median shown, n ≥ 3 mice per condition. Unless otherwise stated: two-tailed unpaired Student’s t tests.
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EpSC proliferation (S-phase EdU labeling) following 24h pharmacological application of PG inhibitors [Indo (Indomethacin; COX non-selective inhibitor), iCOX-1 (SC-560; selective COX-1 inhibitor), iEP4 (ONO-AE3-208; EP4 receptor antagonist)] or PG enhancers [i15-PGDH (SW033291; 15-PGDH inhibitor), dmPGE 2 ]. One-Way ANOVA with Šidák multiple-comparison correction. (C) Experimental schematic for genetic perturbation of LCs. (D-E) Representative IF image and COX-1 MFI of LCs from LV expressing gCtrl or gPtgs1. (F) EpSC proliferation (S-phase EdU labeling) and epidermal thickness in gCtrl or gPtgs1 animals. (G) Experimental schematic of in utero LV delivery for genetic perturbation of EpSCs. (H) Volcano plot of differential gene expression between gCtrl or gPtger4 EpSCs. Genes associated with proliferation, differentiation and barrier function highlighted. (I) Heatmap showing relative Z-score normalized expression of key genes related to epidermal stress and proliferation in gPtger4 or gCtrl EpSCs. (J) Pathway-level analysis using Reactome database in upregulated genes in gPtger4 EpSCs. (K) Representative IF images and quantification of EpSC proliferation (S-phase EdU labeling) in gCtrl or gPtger4 LV + regions. (L) Representative H&E staining and quantifications of epidermal thickness in gCtrl or gPtger4 (transduced with 70% efficiency). For (A-L): Each dot corresponds to data from one mouse with median shown, n ≥ 3 mice per condition. Unless otherwise stated: two-tailed unpaired Student’s t tests.

Journal: bioRxiv

Article Title: A commensally regulated immune rheostat fine-tunes skin barrier fitness

doi: 10.64898/2026.01.07.698149

Figure Lengend Snippet: EpSC proliferation (S-phase EdU labeling) following 24h pharmacological application of PG inhibitors [Indo (Indomethacin; COX non-selective inhibitor), iCOX-1 (SC-560; selective COX-1 inhibitor), iEP4 (ONO-AE3-208; EP4 receptor antagonist)] or PG enhancers [i15-PGDH (SW033291; 15-PGDH inhibitor), dmPGE 2 ]. One-Way ANOVA with Šidák multiple-comparison correction. (C) Experimental schematic for genetic perturbation of LCs. (D-E) Representative IF image and COX-1 MFI of LCs from LV expressing gCtrl or gPtgs1. (F) EpSC proliferation (S-phase EdU labeling) and epidermal thickness in gCtrl or gPtgs1 animals. (G) Experimental schematic of in utero LV delivery for genetic perturbation of EpSCs. (H) Volcano plot of differential gene expression between gCtrl or gPtger4 EpSCs. Genes associated with proliferation, differentiation and barrier function highlighted. (I) Heatmap showing relative Z-score normalized expression of key genes related to epidermal stress and proliferation in gPtger4 or gCtrl EpSCs. (J) Pathway-level analysis using Reactome database in upregulated genes in gPtger4 EpSCs. (K) Representative IF images and quantification of EpSC proliferation (S-phase EdU labeling) in gCtrl or gPtger4 LV + regions. (L) Representative H&E staining and quantifications of epidermal thickness in gCtrl or gPtger4 (transduced with 70% efficiency). For (A-L): Each dot corresponds to data from one mouse with median shown, n ≥ 3 mice per condition. Unless otherwise stated: two-tailed unpaired Student’s t tests.

Article Snippet: The following pharmacological inhibitors were topically applied for 24hrs, all suspended in EtOH at 1mg/ml concentration: Indomethacin (S1723), highly selective COX-1 inhibitor SC-560 (S6686), EP4-receptor antagonist ONO-AE3-208 (E2986), 15-PGDH inhibitor SW033291 (S7900) all from Selleckchem, and dmPGE 2 (Cayman chemicals, Cat: 14750).

Techniques: Labeling, Comparison, Expressing, In Utero, Gene Expression, Staining, Transduction, Two Tailed Test