Panel A shows histograms with the x-axis represents OSMR expression levels, and the y-axis represents cell count. Panel B is a bar graph quantifying extracellular OSMR expression, with the y-axis showing OSMR protein expression (MFI fold change relative to FMO). Panel C is a bar graph quantifying total OSMR expression, with the y-axis showing OSMR protein expression (MFI fold change relative to FMO). Panel D shows immunoblots for pSTAT1 and pSTAT3 in primary fibroblasts from Patient 2 transduced with WT OSMR or an empty vector and a healthy control, before and after treatment with OSM. The x-axis lists the conditions, and the y-axis lists the proteins. Panel E is a bar graph quantifying pSTAT1 signal, with the y-axis showing normalized pSTAT1 signal after OSM stimulation. Panel F is a bar graph quantifying pSTAT3 signal, with the y-axis showing normalized pSTAT3 signal after OSM stimulation. Panel G is a heatmap of differentially expressed genes comparing Patient 2 fibroblasts transduced with WT OSMR or an empty vector against three healthy controls before and after OSM stimulation. The x-axis lists the conditions, and the y-axis lists the genes. Panel H is a Venn diagram showing the overlap of significantly upregulated genes upon OSM stimulation in healthy control cells and Patient 2 rescued cells. Panel I is a heatmap of gene set enrichment analysis (GSEA) of significantly enriched immune pathways in Patient 2 fibroblasts transduced with WT OSMR or an empty vector against three healthy controls before and after OSM stimulation. The x-axis lists the conditions, and the y-axis lists the pathways.
WT OSMR restores the OSM/OSMRβ axis and STAT signaling in primary fibroblasts. (A) Extracellular and total OSMRβ expression were quantified in primary fibroblasts from P2 that were transduced with WT OSMR (rescued) or an EV and compared with primary fibroblasts from an HC using flow cytometry; n = 3. (B) Quantification of A, extracellular OSMRβ expression; n = 3. (C) Quantification of A, total OSMRβ expression; n = 3. (D) Immunoblot in primary fibroblasts from P2 that were transduced with WT OSMR or an EV and compared to primary fibroblasts from an HC for pSTAT1 and pSTAT3 before and after treatment with OSM (100 ng/ml for 15 min); n = 4. (E and F) Quantification of the immunoblot in D for (E) pSTAT1 and (F) pSTAT3. (G) Heatmap signatures of differentially expressed genes comparing P2 fibroblasts (either transduced with WT OSMR or an EV) against three HCs before and after stimulation with OSM (100 ng/ml for 15 min). (H) Overlap in significantly upregulated genes upon OSM stimulation in HC cells (blue) and P2 rescued cells (green) as shown through a Venn diagram. (I) GSEA of significantly enriched immune pathways from the MSigDB Hallmark in P2 (either transduced with WT OSMR or an EV) against three HCs before and after stimulation with OSM (100 ng/ml for 15 min). Heatmap is normalized across the rows and shown as relative expression of sample level enrichment scores. Statistical comparisons based on the Kruskal–Wallis H test. Asterisks denote P values: *< 0.05, **< 0.01. Source data are available for this figure: SourceData F7.