recombinant human fc linked notch ligands  (R&D Systems)

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