human dll 1  (Sino Biological)

Journal: eLife

Article Title: Notch signaling functions in noncanonical juxtacrine manner in platelets to amplify thrombogenicity

doi: 10.7554/eLife.79590

Figure Lengend Snippet: ( A and C ) Histograms showing binding of PAC-1 ( A ) and anti-P-selectin antibody ( C ) to platelets pre-incubated with either DAPT (10 µM) or vehicle for 10 min at RT followed by treatment with either DLL-4 (15 µg/ml) or DLL-1 (15 µg/ml) for 10 min, or with thrombin (Thr, 1 U/ml) for 5 min at 37 °C as indicated. ( B and D ) Ccorresponding mean fluorescence intensities of PAC-1 (n=12) and anti-P-selectin antibody (n=9) binding to platelets, respectively. ( E ) Bar diagram representing ATP secretion from platelet dense granules pre-incubated with either DAPT (10 µM) or vehicle for 10 min at RT followed by treatment with DLL-4 for 10 min (n=4). ( F ) Fura-2-loaded platelets were pre-treated for 5 min either with calcium (1 mM) or EGTA (1 mM) followed by incubation with DLL-4 (15 µg/ml) for 15 min and intracellular Ca 2+ was measured. ( G ) Corresponding bar diagram representing mean concentration of intracellular Ca 2+ over 300 sec of measurement (n=5). ( H ) Platelets were pre-treated with either DAPT (10 µM) or DBZ (10 µM) or vehicle for 10 min at RT followed by treatment with DLL-4 (15 µg/ml) for 10 min at RT. PEVs were isolated and analyzed with Nanoparticle Tracking Analyzer (n=8). ( I ) Immunoblot showing profile of tyrosine phosphorylated proteins in platelets pre-treated with either DAPT (10 µM) or DBZ (10 µM) or vehicle for 10 min at RT followed by treatment with either DLL-4 (15 µg/ml) for 10 min at RT or DLL-1 (15 µg/ml) for 10 min at RT or with thrombin (1 U/ml) for 5 min at 37 °C as indicated (n=4). Arrows indicate position of peptides whose intensity increased in presence of DLL-4. ( J and L ) Immunoblots showing expression of p-PI3K and pAKT in DLL-4 (15 µg/ml for 10 min)-treated platelets in absence or presence of either DAPT (10 µM) or vehicle. ( K and M ) Corresponding densitometric analyses normalised with PI3K (n=3) and AKT (n=7), respectively. Data are presented as mean ± SEM of at least three different experiments. Results were analyzed by RM one-way ANOVA with either Dunnett’s multiple comparisons test ( E , H , K and M ) or Sidak’s multiple comparisons test ( B , D and G ). Figure 3—source data 1. Excel sheet shows numerical data of . Figure 3—source data 2. Unedited and labelled blots of . Figure 3—source data 3. Unedited and unlabelled blots of .

Article Snippet: Peptide, recombinant protein , DLL-1 , Sino Biological , 11635-H08H , 15 µg/ml.

Techniques: Binding Assay, Incubation, Fluorescence, Concentration Assay, Isolation, Western Blot, Expressing

Journal: eLife

Article Title: Notch signaling functions in noncanonical juxtacrine manner in platelets to amplify thrombogenicity

doi: 10.7554/eLife.79590

Figure Lengend Snippet: ( A ) Histogram showing binding of Alexa Fluor 488-labelled fibrinogen (10 µg/ml) to platelets pre-incubated with either DAPT (10 µM) or vehicle for 10 min at RT followed by treatment with either DLL-4 (15 µg/ml) or DLL-1 (15 µg/ml) for 10 min, or with thrombin (1 U/ml) for 5 min at 37 °C as indicated. ( B ) Corresponding mean fluorescence intensity of fibrinogen binding to platelets presented as mean ± SEM. Data are representative of nine (n=9) different experiments-and analysed by RM one-way ANOVA with Sidak’s multiple comparisons test. Figure 3—figure supplement 1—source data 1. Excel sheet shows numerical data of .

Article Snippet: Peptide, recombinant protein , DLL-1 , Sino Biological , 11635-H08H , 15 µg/ml.

Techniques: Binding Assay, Incubation, Fluorescence

Journal: PLoS ONE

Article Title: Jagged-1 Signaling in the Bone Marrow Microenvironment Promotes Endothelial Progenitor Cell Expansion and Commitment of CD133 + Human Cord Blood Cells for Postnatal Vasculogenesis

doi: 10.1371/journal.pone.0166660

Figure Lengend Snippet: (A) Flow cytometric analysis of NGFR expression in control vector- (C), human Jagged-1 (J1)- and human Dll-1 (D1)-transfected HESS-5 cells, and the sorting gate (R2) for NGFR + transfected cells. (B) Western blot analysis of hJagged-1 and hDll-1 proteins in HESS-5 cells. Actin was used as an internal control. (C) Notch1 and Notch2 reporter cell lines transfected with RBP-Jk-luc were co-cultured with HESS-5 stromal cells. Luciferase activity (relative light units) was normalized to the activity of Renilla luciferase. Data are expressed as means ± standard deviation (SD) (n = 3). *P < 0.01 between the indicated values.

Article Snippet: The proteins were transferred to nitrocellulose membranes (Hybond-ECL; Amersham Pharmacia Biotech, Piscataway, NJ), and incubated overnight at 4°C with primary antibodies: goat polyclonal IgG against human Jagged-1 (C-20) (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal IgG against human Dll-1 (H-265) (Santa Cruz Biotechnology), or rabbit polyclonal IgG against actin (Sigma).

Techniques: Expressing, Plasmid Preparation, Transfection, Western Blot, Cell Culture, Luciferase, Activity Assay, Standard Deviation