ligand for egfr  (R&D Systems)

Journal: bioRxiv

Article Title: Extracellular signalling regulates gastrin transcription through site-specific phosphorylation and nuclear redistribution of Menin

doi: 10.64898/2026.04.07.717082

Figure Lengend Snippet: (A) Hematoxylin and eosin (H&E) staining of normal duodenum containing Brunner’s glands (nDUO-BG) and duodenal neuroendocrine tumor (DNET). Dashed boxes indicate regions shown at higher magnification. (B) Immunohistochemical staining for synaptophysin (SYP) confirming neuroendocrine differentiation in DNET. (C-D) Immunohistochemical staining for TGFα and EREG in tumor-associated Brunner’s glands (tBG) and DNET. Dashed boxes indicate tumor-gland interfaces. (E-F) Quantification of TGFα and EREG expression by H-score in nDUO-BG, tBG, and DNET. Data are mean ± SEM; ns, not significant; ****P < 0.0001. (G) EGFR immunostaining in nDUO-BG and DNET showing heterogeneous expression across tissues. (H) Menin immunostaining in nDUO-BG and DNET. (I) Representative FFPE DNET specimens showing cytoplasmic or near-absent Menin expression, accompanied by strong TGFα and EREG staining within tumor cells. (J) Quantification of Menin nuclear-to-cytoplasmic (N/C) ratio in nDUO-BG and DNET. Data are mean ± SEM; ****P < 0.0001. Images were taken at 100X, 200X and 400X. Scale bars: 100 μm (low magnification) and 50 μm (high magnification).

Article Snippet: After 24 h, cells were serum-starved for 24 h, followed by treatment with either recombinant human eregulin (EREG) protein (10 nM), a potent ligand for EGFR (R&D systems, #1195-EP) or 10 μM Forskolin (FSK, ThermoFisher, #66575-29-9), activator of adenylyl cyclase and cyclic AMP or and 10nM phorbol 12-myristate 13-acetate (TPA; Sigma-Aldrich, #P8139) for 4-8 h. Cells were lysed, and luciferase activity was measured using the Dual-Luciferase Reporter Assay System (Promega, #E1980) according to the manufacturer’s instructions.

Techniques: Staining, Immunohistochemical staining, Expressing, Immunostaining

Journal: bioRxiv

Article Title: Extracellular signalling regulates gastrin transcription through site-specific phosphorylation and nuclear redistribution of Menin

doi: 10.64898/2026.04.07.717082

Figure Lengend Snippet: (A) Multiple sequence alignment of the Menin C-terminal region from the indicated vertebrate species showing strong conservation of a basic residue–rich motif encompassing Ser487. Conserved basic residues and Ser487 are highlighted. (B) Schematic of human Menin illustrating the position of Ser487 within NLS1. The expanded sequence highlights Ser487 and surrounding basic residues; constructs used in this study. (C) Immunoblot analysis of AGS cells expressing FLAG-tagged wild-type Menin or Ser487 mutants (S487A, S487D) following treatment with EREG, FSK, or TPA. Whole-cell lysates were probed with antibodies against phospho-Ser487 Menin, FLAG-Menin, and GAPDH. (D, E) Immunoblot analysis of MKN-45G and KATO III cells expressing wild-type Menin following stimulation with EREG, FSK, or TPA. Blots were probed for phospho-Ser487 Menin, FLAG-Menin, and β-tubulin. (F, H) Quantification of phospho-Ser487 Menin in AGS, KATO III and MKN-45G cells. (I) Immunoblot analysis of AGS cells examining activation of cAMP and EGFR downstream kinases under the indicated conditions. (J) Densitometric quantification of signalling outputs shown in (I), expressed as fold change relative to vector control. (K) Time-course of Ser487 phosphorylation in AGS cells stimulated with TPA in the presence of kinase inhibitors; MEK inhibitor (U0126), AKT inhibitor (MK-2206), PKC inhibitor (Gö6983), or combined MEK+AKT inhibition. (L) Quantification of Ser487 phosphorylation kinetics following TPA stimulation with the indicated inhibitors. (M) Area-under-the-curve (AUC) analysis of phosphorylation in (L). Data are presented as mean ± SEM; individual data points represent independent biological replicates (n = 3). Statistical significance was determined by one-way ANOVA with Tukey’s multiple-comparison test (*P < 0.05; **P < 0.01; ****P < 0.0001; ns, not significant).

Article Snippet: After 24 h, cells were serum-starved for 24 h, followed by treatment with either recombinant human eregulin (EREG) protein (10 nM), a potent ligand for EGFR (R&D systems, #1195-EP) or 10 μM Forskolin (FSK, ThermoFisher, #66575-29-9), activator of adenylyl cyclase and cyclic AMP or and 10nM phorbol 12-myristate 13-acetate (TPA; Sigma-Aldrich, #P8139) for 4-8 h. Cells were lysed, and luciferase activity was measured using the Dual-Luciferase Reporter Assay System (Promega, #E1980) according to the manufacturer’s instructions.

Techniques: Sequencing, Residue, Construct, Western Blot, Expressing, Activation Assay, Plasmid Preparation, Control, Phospho-proteomics, Inhibition, Comparison

Journal: Nature Communications

Article Title: IL-17RA signaling promotes the dedifferentiation of Paneth progenitors through ADAM17 to regenerate gut epithelium post-irradiation

doi: 10.1038/s41467-026-70479-z

Figure Lengend Snippet: A Representative images of hematoxylin and eosin staining of the terminal ilea from wild type and Il17ra KO mice 5 days after irradiation (scale bar = 100 μm) (left). Arrows represent areas of damage. Quantification of crypt lengths (right). Mice were 13 and 14 weeks old. N = 6. B Splenic and hepatic bacterial dissemination in wild-type and Il17ra KO mice 5 days after irradiation. Mice were 13 and 14 weeks old. N = 14. C Amounts of lipocalin in the cecal content of irradiated wild-type and Il17ra KO mice measured via ELISA. Mice were 13 and 14 weeks old. N = 14. D Representative immunofluorescent images depicting PCNA (green) and DAPI (blue) from the terminal ilea of wild type and Il17ra KO mice 5 days after irradiation (scale bar = 50 μm) (left) and quantification (right). Mice were 13 and 14 weeks old. N = 6. E Schematic of bone marrow transplantation and irradiation procedure. F Splenic and hepatic bacterial dissemination in bone marrow transferred wild-type and Il17ra KO mice 5 days after irradiation. Mice were 8 and 11 weeks old. N = 9. G Representative images of hematoxylin and eosin staining of terminal ilea from Il17ra fl/fl ;Villin-cre + /− mice 5 days after irradiation (scale bar = 100 μm) (left). Arrows represent areas of damage. Quantification of crypt lengths (right). Mice were 7.2, 7.4, 10 and 14.6 weeks old. N = 21. H Splenic and hepatic bacterial dissemination in Il17ra fl/fl ;Villin-cre + /− mice 5 days after irradiation. Mice were 7.2, 7.9, and 14.6 weeks old. N = 23. I Amounts of lipocalin in the cecal content of irradiated Il17ra fl/fl ;Villin-cre + /− mice measured via ELISA. Mice were 7.2, 7.9, and 14.6 weeks old. N = 19. J Representative immunofluorescent images depicting PCNA (green) and DAPI (blue) from the terminal ilea of Il17ra fl/fl ;Villin-cre + /− mice 5 days after irradiation (scale bar = 50 μm) (left) and quantification (right). Mice were 7.9, and 14.6 weeks old. N = 10. N = number of biologically independent mice. All figures are generated from 3 experiments. Data in Fig. 1A, D was analyzed via Students t test. Data are presented as ± SEM on relevant graphs. (Mann-Whitney test, Two-tailed and Student's T test).

Article Snippet: Equivalent amounts of protein was the utilized for experimentation with the Mouse Epiregulin ELISA Kit (RayBiotech) as per the manufacture’s protocol.

Techniques: Staining, Irradiation, Enzyme-linked Immunosorbent Assay, Transplantation Assay, Generated, MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: IL-17RA signaling promotes the dedifferentiation of Paneth progenitors through ADAM17 to regenerate gut epithelium post-irradiation

doi: 10.1038/s41467-026-70479-z

Figure Lengend Snippet: A Representative images of hematoxylin and eosin staining of terminal ilea in Il17ra fl/fl ;Defa6-cre + /− mice 3 days after irradiation (scale bar = 100 μm) (left). Arrows represent areas of damage. Quantification of crypt lengths (right). Mice were 6.9 and 10.6 weeks old. N = 10. B Splenic and hepatic bacterial dissemination in Il17ra fl/fl ;Defa6-cre + /− mice 5 days after irradiation. Mice were 8.0, 8.1 and 10.6 weeks old. N = 16. C Amounts of lipocalin in cecal content of 5 day post irradiated Il17ra fl/fl ;Defa6-cre + /− mice measured via ELISA. Mice were 8.1 and 10.4 weeks old. N = 10. D Representative immunofluorescent images depicting PCNA (green) and DAPI (blue) from the ileum of Il17ra fl/fl ;Defa6-cre + /− mice 3 days after irradiation (scale bar = 50 μm) (left) and quantification (right). Mice were 6.9, 8.9 and 10.6 weeks old. N = 9. E Representative immunofluorescent images depicting OLFM4 (red) and DAPI (blue) from the ileum of non- irradiated (top) and 5 days post-irradiated (bottom) Il17ra fl/fl ;Defa6-cre + /- mice (scale bar = 50 μm) (left) and quantification (right). Mice were 6.7, 10, and 11.6 weeks old. E = 8. F – I Phylum, family, genus, and species beta diversity between non-irradiated Il17ra fl/fl ;Defa6-cre + /− mice. Mice were 6.4 weeks old. N = 16. J Phylogenetic diversity between non-irradiated and 5 day post irradiated Il17ra fl/fl ;Defa6-cre + /− mice. Mice were 6.4 and 7.1 weeks old. Box plot defined as follows: centre: median, lower bound: 25 th percentile, upper bound: 75 th percentile, lower whisker: extends to minima, upper whisker: extends to maxima. N = 16. K Beta diversity between non-irradiated and 5 day post irradiated Il17ra fl/fl ;Defa6-cre + /- mice. Mice were 6.4 and 7.1 weeks old. N = 16. L , M Terminal ileum cecal contents of cohoused littermate, irradiated, Il17ra fl/fl ;Defa6-cre + /− mice were analyzed for commensal diversity at the phyla (top) and selected family (bottom) levels via 16 S microbial sequencing. Mice were 7.1 weeks old. N = 16. N = number of biologically independent mice. Figure 3A, C, E data is generated from 2 independent experiments. Figure 3B, D data are generated from 3 independent experiments. Data are presented as ± SEM on relevant graphs. (Mann-Whitney test, Two-tailed).

Article Snippet: Equivalent amounts of protein was the utilized for experimentation with the Mouse Epiregulin ELISA Kit (RayBiotech) as per the manufacture’s protocol.

Techniques: Staining, Irradiation, Enzyme-linked Immunosorbent Assay, Whisker Assay, Sequencing, Generated, MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: IL-17RA signaling promotes the dedifferentiation of Paneth progenitors through ADAM17 to regenerate gut epithelium post-irradiation

doi: 10.1038/s41467-026-70479-z

Figure Lengend Snippet: A Gene expression from irradiated Il17ra fl/fl ;Defa6-cre + /− mice (6.7, 8.1 weeks old); each point represents an independent mouse. B Experimental schematic. C Adam17 expression in sorted irradiated Paneth cells after 4 h of IL-17A treatment from Defa6-cre;mT/mG organoids. N = 6. D Gene expression from irradiated Il17ra fl/fl ;Defa6-cre + /− mice (6.7, 8.1 weeks old). N = 13. E Single-cell RNA sequencing of Hif1a related genes from non-irradiated and irradiated Paneth cells (PMID: 29887318). F Terminal ileal Nox1 expression from irradiated Il17ra fl/fl ;Defa6-cre + /− mice (6.7 < 8.1 weeks old). N = 15. G Nox1 expression in sorted irradiated Paneth cells after 4 hours of IL-17A treatment. N = 5. H Single-cell RNA sequencing expression of Adam17 related genes from non irradiated and irradiated Paneth cells (PMID: 29887318). I ADAM17 activity +/- 4 mM H 2 O 2 . J Experimental schematic. K Ileum of irradiated Defa6-cre;mT/mG mice given water or 4% NAC water (GFP = green) (DAPI = blue) (scale bar = 50 μm). GFP + clones = 4 cells in a row. Mice were 8.3 and 13.4 weeks old. N = 9. L Experimental schematic. M Hematoxylin and eosin staining of terminal ilea in Il17ra fl/fl ;Defa6-cre + mice treated with Ad-Y5 or Ad-Adam17 (AdAD17) 5 days after irradiation (scale bar = 100 μm). Mice were 8.9, 14.1 and 15.1 weeks old. N = 11. N Ileum of Il17ra fl/fl ;Defa6-cre + mice treated with Ad-Y5 or AdAD17 5 days after irradiation (PCNA = green) (DAPI = blue) (scale bar = 50 μm). Mice were 8.9, 14.1 and 15.1 weeks old. N = 9. O Bacterial dissemination in Il17ra fl/fl ;Defa6-cre + /− mice treated with Ad-Y5 or AdAD17 5 days after irradiation. Mice were 8.9, 14.1 and 15.1 weeks old. N = 14. P EREG in the ileum of Il17ra fl/fl ;Defa6-cre + /− mice treated with Ad-Y5 or AdAD17 5 days after irradiation. Mice were 8.9, 14.1 and 15.1 weeks old. N = 20. N = number of biologically independent mice. 6 K, 6 M, 6 N, 6 O = 2 independent experiments. 6 A, 6 C, 6D, 6 F, 6 G, 6 L = 3 independent experiments. Outliers removed via GraphPad Prism (ROUT analysis, Q = 1%) in Fig. 6A. Data are presented as ± SEM on relevant graphs (Mann-Whitney test, Two-tailed and Wilcox paired test, Two-tailed).

Article Snippet: Equivalent amounts of protein was the utilized for experimentation with the Mouse Epiregulin ELISA Kit (RayBiotech) as per the manufacture’s protocol.

Techniques: Gene Expression, Irradiation, Expressing, Single Cell, RNA Sequencing, Activity Assay, Clone Assay, Staining, MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: IL-17RA signaling promotes the dedifferentiation of Paneth progenitors through ADAM17 to regenerate gut epithelium post-irradiation

doi: 10.1038/s41467-026-70479-z

Figure Lengend Snippet: A Representative images of hematoxylin and eosin staining of terminal ilea from Adam17 fl/fl ;Defa6-cre + /− mice 5 days post irradiation (scale bar = 100 μm) (left). Quantification of crypt lengths (right). Mice were 14.9 and 16 weeks old. N = 10. B Amounts of lipocalin in the cecal content of 5 day post irradiated Adam17 fl/fl ;Defa6-cre + /− mice measured via ELISA. Mice were 6.9, 7 and 16 weeks old. N = 17. C Splenic and hepatic bacterial dissemination from Adam17 fl/fl ;Defa6-cre + /− mice 5 days post irradiation. Mice were 6.9, 7 and 16 weeks old. N = 21. D Representative immunofluorescent images depicting PCNA (green) and DAPI (blue) from the ileum of Adam17 fl/fl ;Defa6-cre + /− mice 5 days after irradiation (scale bar = 50 μm) (left) and quantification (right). Mice were 8.4, 14.9 and 16 weeks old. N = 10. E Representative immunofluorescent images depicting Lyz1 (green), PCNA (red) and DAPI (blue) from the ileum of Adam17 fl/fl ;Defa6-cre + / − mice 5 days after irradiation (scale bar = 50 μm) (left) and quantification (right). Mice were 14.9 and 16 weeks old. N = 10. F RT-qPCR analysis depicting Dll1, Dll4, Anxa1, Anxa3, Anxa5, Nox1, Adam10, Wnt9b, Atoh1, Olfm4, Ly6a , and Ly6d expression in the terminal ileum from Adam17 fl/fl ;Defa6-cre + /− mice 5 days post irradiation. Mice were 6.9 and 7 weeks old. N = 11. G Representative immunohistochemistry images depicting NOX1 from the ileum of Adam17 fl/fl ;Defa6-cre + /− mice 5 days after irradiation (scale bar = 50 μm) (left) and quantification (right). Mice were 14.9, and 16 weeks old. N = 9. H Representative immunofluorescent images depicting ZO-1 (green) and DAPI (blue) from the ileum Adam17 fl/fl ;Defa6-cre + /− mice 5 days after irradiation (scale bar = 50 μm) (left). Quantification via mean fluorescence intensity (right). N = 8. I RT-qPCR results from Adam17 fl/fl ;Defa6-cre + /− mice 5 days post irradiation depicting Ocln and Cldn4 . Mice were 6.9 and 7 weeks old. N = 12. N = number of biologically independent mice. All experiments are representative of 3 experiments. Data are presented as ± SEM on relevant graphs. (Mann-Whitney test, Two-tailed).

Article Snippet: Equivalent amounts of protein was the utilized for experimentation with the Mouse Epiregulin ELISA Kit (RayBiotech) as per the manufacture’s protocol.

Techniques: Staining, Irradiation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Immunohistochemistry, Fluorescence, MANN-WHITNEY, Two Tailed Test

Journal: Biomolecules and Biomedicine

Article Title: ERBB4 as a therapeutic target in aortic dissection: Implications for cell-based therapies in vascular regeneration

doi: 10.17305/bb.2025.11925

Figure Lengend Snippet: Phenotype transition of VSMC cells. (A) IHC results revealed the decreased levels of α-actin, and TAGLN in aortic tissue; (B) Western blot results presented the decreased levels of α-actin, SPP1, TAGLN, and the highly expressed of epiregulin in aortic tissue. Bar graphs presented the quantification of α-actin (C), SPP1 (D), TAGLN (E), and epiregulin (F). vs Control group, ** P < 0.01, *** P < 0.001. VSMC: Vascular smooth muscle cell; IHC: Immunohistochemical; TAGLN: Transgelin.

Article Snippet: To prevent non-specific binding, membranes were blocked with 5% non-fat milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 h at room temperature, followed by overnight incubation at 4 ∘ C with primary antibodies against ERBB4 (1:1000, Proteintech, Wuhan, China), α-actin (1:1000, Proteintech), Transgelin (TAGLN) (1:1000, Elabsience, Shanghai, China), SPP1 (1:1000, ZENBIO, Chengdu, China), epiregulin (1:1000, Biorbyt, UK), CD151 (1:1000, Proteintech), ITGAE (1:1000, ab224202, Abcam, UK), ITGB5 (1:1000, Proteintech), p-IκBα (1:5000, Proteintech), IκBα (1:5000, Proteintech), p-NF-κB-65 (1:2000, Proteintech), NF-κB-65 (1:1000, Proteintech), and GAPDH (1:50000, Proteintech) as a loading control.

Techniques: Western Blot, Control, Immunohistochemical staining

Journal: bioRxiv

Article Title: Mapping mesenchymal diversity in the developing human intestine and organoids

doi: 10.1101/2025.07.22.665939

Figure Lengend Snippet: UMAP projections of the major cell class composition in a Xenium image of both an A) EGF-tHIO and B) EREG-tHIO. C) Stacked bar graph summarizing the percent cellular composition of the EGF-tHIO (left) and EREG-tHIO (right) samples. Cell ID masks generated in XE depicting 1) the entire sample acquired for the D) EGF-tHIO and E) EREG-tHIO. Scale bar = 5000µm. The white box indicates the ROI depicted in D2/E2-D7/E7. D2-7 and E2-7 display the cell sub-type distribution of each major cell class identified using the Xenium panel visualized as 2) the major cell class mask, 3) the epithelium sub-types, 4) the fibroblast sub-types, 5) the SMC-related sub-types, 6) the ENS sub-types, and 7) the endothelium sub-types. Scale bar = 500µm Cell classes are color matched in A-E ; epithelium (gold), fibroblasts (navy), muscularis mucosa (MM, light green), SMC and pericytes (dark green), ENS (yellow), and endothelium (cyan). No immune cells were observed in either tHIO sample. DAPI staining of cell nuclei depicted in grey.

Article Snippet: Organoid basal growth media was supplemented with epidermal growth factor (EGF) (100 ng/mL R&D Systems Cat#236-EG-01M) or Epiregulin (EREG) (10 ng/mL R&D Systems Cat#1195-EP-025/CF) with Noggin-Fc (100ng/mL, purified from conditioned media), and R-Spondin1 (5% conditioned medium) for the first three days of culture to pattern a proximal small intestine.

Techniques: Generated, Staining

Journal: bioRxiv

Article Title: Mapping mesenchymal diversity in the developing human intestine and organoids

doi: 10.1101/2025.07.22.665939

Figure Lengend Snippet: UMAP projections of the fibroblast cell class composition in a Xenium image of both an A) EGF-tHIO and B) EREG-tHIO. C) Stacked bar graph summarizing the percent cellular composition of the EGF-tHIO (left) and EREG-tHIO (right) samples. Cell ID masks generated in XE depicting D) EGF-tHIO and E) EREG-tHIO samples. 1) XE cell mask summarizing the major cell classes for the tHIOs to landmark fibroblast population locations. Fibroblast sub-type distribution is depicted as 2) the F3 + SECs, 3) the FABP5 + early LPFs, 4) the ADAMDEC1 + late LPFs, 5) the SHISA3 + SMFs, and 6) the CXCL13 + Fibroblasts. 7) depicts the composite mask summarizing all fibroblast cell types in the tHIOs. Scale bars = 100µm

Article Snippet: Organoid basal growth media was supplemented with epidermal growth factor (EGF) (100 ng/mL R&D Systems Cat#236-EG-01M) or Epiregulin (EREG) (10 ng/mL R&D Systems Cat#1195-EP-025/CF) with Noggin-Fc (100ng/mL, purified from conditioned media), and R-Spondin1 (5% conditioned medium) for the first three days of culture to pattern a proximal small intestine.

Techniques: Generated