notch ligand dll4  (R&D Systems)

Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a) αSMA mRNA levels in HCAEC treated with TGFβ1 (10 ng/ml) or exposed to hypoxic conditions (5% O 2 ), in the absence or presence of moderate dose EtOH (25 mM, green bars) or high dose EtOH (100 mM, red bars) +/- the γ-secretase inhibitor DAPT (20 μM). (b) αSMA and fibronectin (FN) mRNA levels in HCAEC treated with TGFβ1 +/- the Notch ligand DLL4 (3 μg/ml) in the absence or presence of DAPT (20 μM). Data are mean±SEM, n=3. *p<0.05 vs control (no treatment), #p<0.05 vs TGFβ1 or hypoxia.

Article Snippet: Where indicated, cells were treated with Notch inhibitor DAPT (20 μM, Cat. #2634, Tocris,) or Notch ligand DLL4 (2 μg/ml, Cat. # 1506-D4, R&D Systems).

Techniques: Control

Journal: Nature Communications

Article Title: Human endothelial cells promote a human neural stem cell type B phenotype via Notch signaling

doi: 10.1038/s41467-025-60194-6

Figure Lengend Snippet: a UMAP demonstrating hEC and hNSPC clusters along with featureplots of Notch ligands DLL4, JAG1, and JAG2 expressed by hECs. b Featureplots showing the expression of Notch receptors NOTCH1, NOTCH2, and NOTCH3 by hNSPCs. c SC27 hNSPCs plated on DLL4 ligand increased the expression of HEY1 (**p = 0.0011), HES4 (*p = 0.0114), GFAP (*p = 0.0123), and SOX2 (*p = 0.0115) relative to control. d DLL4 expression was reduced in hECs treated with DLL4 siRNA compared to non-targeting (NT) control (**** p < 0.0001). e SC27 hNSPCs in MC and co-culture with hECs treated with NT siRNA or DLL4 siRNA at day 1 after plating were stained for GFAP, SOX2, and CD31. f The percentage of GFAP + SOX2+ (**** p < 0.0001) and GFAP+ (**** p < 0.0001) hNSPCs significantly increased in NT siRNA-treated hEC co-cultures compared to MC. DLL4 siRNA-treated hECs significantly decreased GFAP + SOX2+ (**** p < 0.0001) and GFAP+ (**** p < 0.0001) hNSPCs compared to NT control. There was no significant difference between MC and DLL4-siRNA treated hEC co-cultures in terms of GFAP+SOX2+ type B hNSPCs (p = 0.4583). GFAP+ cells significantly increased in DLL4 siRNA-treated hEC co-cultures compared to MC (**p = 0.0079). SOX2+ cells significantly decreased in DLL4 siRNA-treated hEC co-cultures compared to MC (**p = 0.0058). Relative expression determined via qRT-PCR normalized to GAPDH ( c ) or RPLP0 ( d ). Nuclei were stained with Hoechst. Scale bars, 100 µm, n = 3 independent biological replicates indicated by symbols, 5 areas quantified per sample, percentages are out of total hNSPCs, mean with SEM. Analysis used ( c , d ) unpaired two-tailed Student’s t-test and ( f ) one-way ANOVA with Tukey post-hoc test for multiple comparisons. Source data are provided as a file.

Article Snippet: Recombinant human Notch ligand Fc chimera proteins DLL4, JAG1, and JAG2 (R&D Systems) were plated on coverslips at a 20 μg/mL concentration along with laminin after PDL coating.

Techniques: Expressing, Control, Co-Culture Assay, Staining, Quantitative RT-PCR, Two Tailed Test

Journal: Nature Communications

Article Title: Human endothelial cells promote a human neural stem cell type B phenotype via Notch signaling

doi: 10.1038/s41467-025-60194-6

Figure Lengend Snippet: a 15-year-old human SVZ was stained for GFAP, CD31, and DAPI and imaged with confocal microscopy (scale bar 50 μm) ( n = 1, biological repeats at other stages in Supplementary Fig. ). Arrowheads point to examples of GFAP+ processes contacting vasculature in the niche. Box denotes region for maximum projection of 3 confocal images (1 µm each, middle panel, scale bar 50 μm) to show overlap between GFAP+ processes and CD31+ vessels, which is shown in greater detail in zoomed panel (right, scale bar 25 μm) and denoted by yellow arrowheads. b 2-year-old human SVZ was stained for GFAP, SOX2, CD31, and DAPI and imaged with confocal microscopy ( n = 1, biological repeats at other stages in Supplementary Fig. ). SOX2+ nucleus surrounded by GFAP is denoted by the fat arrow, while GFAP+ process extending from the cell is marked by the thin arrow. The GFAP+ process contacts the CD31+ vessel (V) along the region denoted by the arrowhead. Scale bar 10 μm. c 6-year-old human SVZ stained for GFAP, PROM1, CD31, and DAPI and imaged with confocal microscopy ( n = 1, biological repeats at other stages in Supplementary Figs. , ). Processes from cells co-expressing GFAP and PROM1 extend toward and wrap around CD31+ vessels. Bottom right panel represents maximum projection of 9 confocal images (1 μm each). Scale bars, 30 um. d Schematic summarizing hEC-hNSPC reciprocal communication. HEC contact increases the percentage of hNSPCs with a type B cell phenotype via Notch signaling involving the Notch ligand DLL4 on hECs and Notch receptor on hNSPCs. As shown previously, hNSPCs increase hEC vessel formation, creating a positive feedback interaction between these critical cell types . Created with BioRender.com.

Article Snippet: Recombinant human Notch ligand Fc chimera proteins DLL4, JAG1, and JAG2 (R&D Systems) were plated on coverslips at a 20 μg/mL concentration along with laminin after PDL coating.

Techniques: Staining, Confocal Microscopy, Expressing

Journal: Science immunology

Article Title: Epiregulin is a dendritic cell-derived EGFR ligand that maintains skin and lung fibrosis

doi: 10.1126/sciimmunol.abq6691

Figure Lengend Snippet: (A) Expression fold change of EREG when THP-1 monocytes were incubated with each indicated cytokine. (B) EREG protein quantification from supernatant of THP-1 incubated with IFNa2 for 4 hours, n=4 per group, data is representative from 2 independent experiments. (C-E) EREG expression fold change from freshly isolated peripheral blood CD14+ monocytes (C) and CD1c+ dendritic cell precursors (D) or cultured human BMDC (E) after incubation with IFNα2. (F) Expression fold change of NOTCH ligands, receptors, and target genes by HFFs incubated with recombinant human EREG (n=5). (G) HES1 expression fold change in SSc fibroblasts after incubation with EREG for 4 hours, n=5 per group. (H) EREG relative expression by BMDC primed with IFNα2 prior to exposure to NOTCH ligand DLL4 (n=3–4 per time point in each group). Statistics compare each group ± DLL4. (I) Relative expression of EGFR ligands by HFF (n=3). Genes with fewer than three points were below detectable level. (J) Changes in ECM gene expression when HFF were incubated with media alone (NT) or EREG neutralizing antibody (Ereg Ab). FNEDA refers to the extra domain A-containing isoform of fibronectin (n=5 per group). (K) Model of EREG-NOTCH circuit between monocyte-derived DC3 and fibroblasts. Data are means ± SD (ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P<0.0001) analyzed with unpaired two-tailed Student t test (A-H, J) and one-way ANOVA with Tukey multiple-comparisons test (I).

Article Snippet: To test the effects of NOTCH ligands, monocytes were incubated in media alone or IFNa2 1000 U/mL for 6 hours at 37° C. During the last 45 minutes, recombinant NOTCH ligands DLL4 (R&D Biosciences 1506-D4) and NOV (R&D Biosciences 1640-NV) each 10 μg/mL were adhered to the bottom of 48 well plates at 37° C for 45 minutes as described ( 90 ).

Techniques: Expressing, Incubation, Isolation, Cell Culture, Recombinant, Gene Expression, Derivative Assay, Two Tailed Test