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: International journal of molecular sciences

Article Title: Endothelial Sprout Formation Is Regulated by Substrate Stiffness and Notch Signaling.

doi: 10.3390/ijms26073155

Figure Lengend Snippet: Figure 5. Expression of Dll4 and NICD in sprouted spheroids after the chemical influence of the Notch signaling pathway. (A–D) Notch ligand (Dll4) and receptor (NICD) intensities (a.u. arbitrary units) in sprouted HUVEC spheroids. Spheroid sprouting was influenced by the addition of the compounds listed above in the medium during spheroid embedding. Intensities were analyzed in the complete spheroid (overall) as well as single cells (tip and stalk cells). (A) Overall Dll4 intensity. Data from three independent experiments, each with at least triplicates, are shown in a scatter plot as mean ± s.d. (B) Dll4 intensities in tip and stalk cells shown in a violin plot as min to max. * p < 0.1; ** p < 0.01; **** p < 0.0001; ns, not significant (one-way ANOVA followed by Šídák’s multiple comparisons test). (C) Overall NICD intensity. Data from three independent experiments, each with at least triplicates, are shown in a scatter plot as mean ± s.d. ** p < 0.01 (one-way ANOVA followed by Tukey’s multiple comparison test). (D) NICD intensities in tip and stalk cells shown in a violin plot as min to max. * p < 0.1; ** p < 0.01; ns, not significant (one-way ANOVA followed by Šídák’s multiple comparisons test). (E,F) Representative images of sprouted spheroids; incubation for 24 h. Cells are stained for nuclei (blue) and Dll4 (red) or NICD (green). Scale bar: 100 µm.

Article Snippet: Recombinant Human DLL4 was obtained from R&D Systems (Wiesbaden, Germany) and used at a working concentration of 1 μg/mL.

Techniques: Expressing, Comparison, Incubation, Staining

Journal: International journal of molecular sciences

Article Title: Endothelial Sprout Formation Is Regulated by Substrate Stiffness and Notch Signaling.

doi: 10.3390/ijms26073155

Figure Lengend Snippet: Figure 6. Expression of Dll4 and NICD in sprouted spheroids after stiffness modification of the surrounding matrix. (A–D) Notch ligand (Dll4) and receptor (NICD) intensities (a.u. arbitrary units) in sprouted HUVEC spheroids. A photoinducible collagen I hydrogel was applied as a surrounding matrix. Spheroid sprouting was influenced by stiffness changes due to different exposure times to UV light as indicated above. Data from three independent experiments, each with at least triplicates, are shown in a scatter plot as mean ± s.d. ** p < 0.01 (one-way ANOVA followed by Tukey’s multiple comparison test). (B) Dll4 intensities in tip and stalk cells shown in a violin plot as min to max. * p < 0.1; ns, not significant (one-way ANOVA followed by Šídák’s multiple comparisons test). (C) Overall NICD intensity. Data from three independent experiments, each with at least triplicates, are shown in a scatter plot as mean ± s.d. * p < 0.1; **** p < 0.0001 (one-way ANOVA followed by Tukey’s multiple comparison test). (D) NICD intensities in tip and stalk cells shown in a violin plot as min to max. * p < 0.1; **** p < 0.0001; ns, not significant (one-way ANOVA followed by Šídák’s multiple comparisons test). (E,F) Representative images of sprouted spheroids; incubation for 24 h. Cells are stained for nuclei (blue) and Dll4 (red) or NICD (green). Scale bar: 100 µm.

Article Snippet: Recombinant Human DLL4 was obtained from R&D Systems (Wiesbaden, Germany) and used at a working concentration of 1 μg/mL.

Techniques: Expressing, Modification, Comparison, Incubation, Staining