Journal: New biotechnology

Article Title: Clinically compatible advances in blood-derived endothelial progenitor cell isolation and reprogramming for translational applications.

doi: 10.1016/j.nbt.2021.02.001

Figure Lengend Snippet: Fig. 1. EPC morphology and gene expression. (A) Appearance of a freshly isolated EPC colony at passage 0 on day 9 of culture. (B) Appearance of EPCs in culture at passage 2, (C–F) EPCs express the endothelial cell specific markers CD31 (C, red, DAPI blue) and CD144 (D, green, DAPI blue) and pluripotency associated markers NANOG (E, red) and REX-1 (F, red, DAPI blue). Images A-B at 50X magnification, images C, D, E, F at 100X magnification.

Article Snippet: Primary EPCs were stained using specific antibodies against CD31 (Cat. no. BBA7, R&D Systems, Abingdon, UK), CD144 (Cat. no. MAB9381, R&D Systems) and REX1 (Cat. no. AF3598, R&D Systems), or by using ULEX (Cat. no. RL-1062-2, Vector Laboratories) or DAPI (Cat. no. H-1200-10, Vector Laboratories) at the dilution recommended by the manufacturer.

Techniques: Gene Expression, Isolation

Journal: Sleep

Article Title: Complement promotes endothelial von Willebrand factor and angiopoietin-2 release in obstructive sleep apnea

doi: 10.1093/sleep/zsaa286

Figure Lengend Snippet: IH promotes co-localization of intracellular CD59 with WPB in ECs. (A) Representative confocal images of DuoLink signal between CD59 and Weibel-Palade Body (WPB) in HUVECs in normoxia and IH. EC plasma membrane is identified by immunofluorescence for VE–cadherin. Scale bar: 10 μm. (B) Quantitation of DuoLink signal in normoxia and IH (n = 4). (C) Western blot probed with antibodies directed against CD59 and vWF in the immunoprecipitate of vWF in HUVECs exposed to normoxia and IH. (D) Western blot probed with antibodies directed against CD59 and STX3 in the immunoprecipitate of STX3 in HUVECs exposed to normoxia and IH. The experiment was reproduced three times. (E) Western blot probed with antibodies directed against Cav1.2, Cav3.1, and STX3 in the immunoprecipitate of STX3 in HUVECs exposed to normoxia and IH. IgG served as negative control in (C), (D), and (E). Data in B are shown as the mean ± SE, two-sided t-test. Cav1.2, Voltage-sensitive Calcium Channel L-type 1.2; Cav3.1, Voltage-sensitive Calcium Channel T-type 3.1; STX3, Syntaxin-3. Other abbreviations as in Figure 2.

Article Snippet: CD59 and MAC in ECs from participants: For assessment of cellular distribution of CD59, harvested ECs were identified by positive staining with goat anti-human polyclonal antibodies directed against VE-cadherin (R&D Systems) followed by FITC-conjugated donkey anti-goat secondary antibodies (Jackson ImmunoResearch).

Techniques: Membrane, Immunofluorescence, Quantitation Assay, Western Blot, Negative Control

Journal: Brain, Behavior, & Immunity - Health

Article Title: CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium

doi: 10.1016/j.bbih.2021.100370

Figure Lengend Snippet: – Decrease in VE-cadherin in response to CCL4. hCMEC/D3 were left untreated or treated with CCL4 (100 ​ng/mL) for 5, 10 and 60 ​min, fixed and stained for VE-cadherin and nuclear DNA. A- Images show full projection of confocal stacks through the full thickness of the cells. Images were taken on Zeiss LSM700. Scale bar 50 ​μm (top row) or 10 ​μm (middle and bottom row). B - Mean area average ​± ​SEM of VE-cadherin stain, expressed as VE-cadherin fold change. Statistical analysis used a One-way ANOVA and Dunnet post hoc test. ∗p ​< ​0.05. P ​= ​0.0462, F ​= ​3.992, and dF ​= ​15. C – Western blot and analysis of total VE-Cadherin. Normalized Mean VE-cadherin ​± ​SEM, normalized to HSC70 is shown as fold change. Statistical analysis used a One-way ANOVA and Dunnet post hoc test.

Article Snippet: Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were performed as previously described ( ) using antibodies against the proteins of interest: CCR5 (Abcam) was used at 1:3,000, HSC70 (Sigma) at 1:10,000 and P–P38 MAPK (T180/Y182) at 1:3000 (Abcam), anti-Human VE-Cadherin Monoclonal Antibody (R&D Systems) at 1:3000 in 0.1% PBSA.

Techniques: Staining, Western Blot

Journal: Brain, Behavior, & Immunity - Health

Article Title: CCL4 induces inflammatory signalling and barrier disruption in the neurovascular endothelium

doi: 10.1016/j.bbih.2021.100370

Figure Lengend Snippet: - CCL4 significantly reduces VE-Cadherin signal from the plasma membrane. VE-cadherin expression following CCL4 treatments for 5-, 10-, 30- and 60-min. Confluent hCMEC/D3 were left untreated or were treated with CCL4 (100 ​ng/mL) for the above mentioned timepoints, stained for the extracellular epitope of VE-cadherin and nuclear DNA. A - Images show full projection of confocal stacks through the full thickness of the cells. Images were taken on Zeiss LSM700. Scale bar 50 ​μm. B- Experimental detail: single endothelial cell and primary (green) and secondary antibodies (grey). C-– Mean area average ​± ​SEM of VE-cadherin stain, normalized to control. Statistical analysis used One-way ANOVA and Dunnet post hoc test. ∗, P ​≤ ​0.05; ∗∗, P ​≤ ​0.01. P ​= ​0.0095, F ​= ​4.512, and dF ​= ​32. D - VE-cadherin expression following CCL4 treatments for 5, 10, 30 and 60 min hCMEC/D3 were left untreated or were treated with CCL4 (100 ​ng/mL) for the above mentioned timepoints, stained for the extracellular epitope of VE-cadherin, intracellular epitope (C-19) and nuclear DNA. VE-cadherin Z stacks were taken on Zeiss LSM700 using a 63x and a 10x objective. A - Images show full projection of confocal stacks through the full thickness of the cells. Images were taken on Zeiss LSM700. Scale bar 20 ​μm (top row only) or 50 ​μm. E − Experimental detail: single endothelial cell, and primary (green and red) and secondary (grey) antibodies. F - JACoP plugin on image J was used to calculate the coefficient between the red/green channels for pixel co-localis ation in 10x images. Statistical analysis performed using a One-way ANOVA and Dunnet post hoc test. ∗, P ​≤ ​0.05. analysis performed using a One-way ANOVA and Dunnet post hoc test. ∗∗∗, P ​< ​0.001. P ​= ​0.0001, F ​= ​28.19, and dF ​= ​11. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were performed as previously described ( ) using antibodies against the proteins of interest: CCR5 (Abcam) was used at 1:3,000, HSC70 (Sigma) at 1:10,000 and P–P38 MAPK (T180/Y182) at 1:3000 (Abcam), anti-Human VE-Cadherin Monoclonal Antibody (R&D Systems) at 1:3000 in 0.1% PBSA.

Techniques: Clinical Proteomics, Membrane, Expressing, Staining, Control

Journal: Cell reports

Article Title: SOX17 integrates HOXA and arterial programs in hemogenic endothelium to drive definitive lympho-myeloid hematopoiesis

doi: 10.1016/j.celrep.2021.108758

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD144 PE-Vio770 (clone: 12G5) , Miltenyi Biotec , Cat# 130-100-720; RRID:AB_2655158.

Techniques: Recombinant, Chromatin Immunoprecipitation, Control, Expressing, Plasmid Preparation, Software