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: 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