Journal: bioRxiv

Article Title: Prostaglandin E₂ Reverses Myofibroblast Differentiation in Eosinophilic Esophagitis

doi: 10.64898/2026.01.27.702012

Figure Lengend Snippet: A. Experimental schematic of bulk RNA-seq analysis of FEF3 stimulated with TGFβ or treated with PGE₂, and pseudo-bulk sequencing of fibroblasts isolated from subjects with active EoE or remission. B. C. D. Top 10 enriched terms in TGFβ stimulated FEF3 and depleted terms in PGE₂ treatment FEF3. And Top 10 enriched terms in fibroblasts from active EoE patients compared with remission EoE patients. Gene Set Enrichment Analysis for Integrin1 pathway in the PID based on each condition. Venn diagrams depicting enrichment gene in PID_ Integrin1 pathway and the number of DEGs. The threshold for DEGs set by log2 fold change –1.5 or 1.5 and p < 0.05. All data sets were analyzed on Gene Set Enrichment Analysis based on the PID. Dot size and color represent the number of core enrichment genes and the normalized enrichment score (NES) for the pathway. FDR, false discovery rate. E. Venn diagrams depicting overlapping genes in each condition ( B, C, and D). F. MA plots based on RNA sequencing data. The x-axis is the mean of the normalized counts and the y-axis is the log2(fold change). DEGs with p < 0.05 and log2(fold change) > 1.5 were plotted in red, and DEGs with p < 0.05 and log2(fold change) <-1.5 were plotted in blue. THBS-1 was plotted as orange.

Article Snippet: The Human THBS-1 Quantikine ELISA Kit (DTSP10; R&D Systems) is a quantification of human THBS-1 in cell culture supernatants.

Techniques: RNA Sequencing, Sequencing, Isolation

Journal: bioRxiv

Article Title: Prostaglandin E₂ Reverses Myofibroblast Differentiation in Eosinophilic Esophagitis

doi: 10.64898/2026.01.27.702012

Figure Lengend Snippet: A. Relative THBS-1 expression by qPCR in FEF3 myofibroblasts treated for 24 hours with PGE₂ (1 μM) (n = 3). B. Relative THBS-1 expression by qPCR in fibroblasts from active and remission EoE patients stimulated with TGFβ (10 ng/mL) with or without PGE₂ for 24 hours (n = 7). C. Relative THBS-1 expression by qPCR in fibroblasts from active and remission EoE patients (n = 7) D. Representative images of immunohistochemistry of THBS-1 in human biopsy tissues. Scale bar 100µm. E. Relative THBS-1 protein in supernatant by ELISA. FEF3 were stimulated with TGFβ and treated with or without PGE₂ for 15 minutes. F. Transfection with siRNA against YAP in FEF3. Scramble siRNA-transfected FEF3 were used as a control. Twenty-four hours after transfection, fibroblasts were stimulated with TGFβ. Relative THBS-1 expression by qPCR (n = 3). G. Relative THBS-1 expression in FEF3 myofibroblasts treated with verteporfin by qPCR (n = 3). H. Transfection with siRNA against THBS-1 in FEF3. Scramble siRNA-transfected FEF3 were used as a control. Twenty-four hours after transfection, fibroblasts were stimulated with TGFβ. Relative THBS-1, αSMA, FN1, and COL1A1 proteins expression by western blot. I. The EdU staining is used to detect cell proliferation. The cell nuclei are stained blue (DAPI), and the EdU+ nuclei are stained red. Quantifying the percentage of EdU+ nuclei in the nucleus per HPFs. For each condition, randomly selected HPFs (n = 10) were analyzed, and the mean value per condition was calculated. FEF3 stimulated with TGFβ and THBS-1 (100 and 1000 ng/mL). Scale bar 100µm. J. The EdU staining is used to detect cell proliferation. Transfection with siRNA against THBS-1 in FEF3. The cell nuclei are stained blue (DAPI), and the EdU+ nuclei are stained red. Quantifying the percentage of EdU+ nuclei in the nucleus per HPFs. For each condition, randomly selected HPFs (n = 10) were analyzed, and the mean value per condition was calculated. Scale bar 100µm. All data were representative of 3 independent experiments and expressed as means ± SDs. A, B, C, D, E, F, and G. 1-way analysis of variance was performed for statistical analyses. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The Human THBS-1 Quantikine ELISA Kit (DTSP10; R&D Systems) is a quantification of human THBS-1 in cell culture supernatants.

Techniques: Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Transfection, Control, Western Blot, Staining

Journal: bioRxiv

Article Title: Prostaglandin E₂ Reverses Myofibroblast Differentiation in Eosinophilic Esophagitis

doi: 10.64898/2026.01.27.702012

Figure Lengend Snippet: Butaprost de-differentiates myofibroblasts and collagen deposition via YAP/THBS-1 axis. A. Schematic of murine EoE model and Butaprost treatment. B. Representative images of hematoxylin and eosin (H&E), trichrome staining, immunohistochemistry for THBS-1 and YAP of the murine esophagus. Scale bar, 50 μm for H&E, THBS-1, and YAP, 100μm for trichrome staining. C. Thickness (μm) of lamina propria and number of nuclear YAP-stained cells per high-power field. Thickness was measured in 10 randomly selected high-power fields (HPFs), and YAP staining was analyzed in five randomly selected HPFs. Data were obtained from four mice per group. Group means were calculated from individual mouse averages. D. Representative flow cytometry histogram of THBS-1, αSMA, and FN1 in the murine esophagi. E. Quantification of THBS-1, αSMA, and FN1 expression in the murine esophagi as measured by flow cytometry (n = 4). Data are indicated as means ± SDs. One-way analysis of variance (C, D, and E) was utilized for statistics. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. ns not significant

Article Snippet: The Human THBS-1 Quantikine ELISA Kit (DTSP10; R&D Systems) is a quantification of human THBS-1 in cell culture supernatants.

Techniques: Staining, Immunohistochemistry, Flow Cytometry, Expressing

Journal: bioRxiv

Article Title: Prostaglandin E₂ Reverses Myofibroblast Differentiation in Eosinophilic Esophagitis

doi: 10.64898/2026.01.27.702012

Figure Lengend Snippet: PGE₂ induces YAP phosphorylation and degradation by proteasome via the cAMP pathway. YAP downregulation induced decreasing αSMA and THBS-1. Furthermore, decreasing THBS-1 induces downregulating ECM proteins such as FN1 and COL1A1 and fibroblast proliferation.

Article Snippet: The Human THBS-1 Quantikine ELISA Kit (DTSP10; R&D Systems) is a quantification of human THBS-1 in cell culture supernatants.

Techniques: Phospho-proteomics