Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: Nogo-B affected HUVECs proliferation and tube formation by transfected with GV248-Nogo-B RNAi-GFP. (A) The cells grew well and shaped like spindles with no vacuole seen in the nucleus and were transfected under a fluorescence microscope and the whole cell emits fluorescence (200X). (B) Detection of the Nogo-B expression by Western Blot. Nogo-B was completely knocked down. (C) The effect of Nogo-B on proliferation and (D) tube-formation of HUVECs were measured by CCk-8 assay and tube formation assay. Data was shown as means ± SD, (C) n=3; (D) n=5; *P<0.05, **P<0.01, ***P<0.001. Scar bars: 100 μm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Transfection, Fluorescence, Microscopy, Expressing, Western Blot, CCK-8 Assay, Tube Formation Assay

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: Cytokines detected in HUVECs supernatant after knocked down Nogo-B. A. Cytokines in supernatant of HUVEC-NC and HUVEC-siNogo-B detected by antibody microarray. They were labeled by Cy3 and Cy5, which were two different fluorescent molecular. B. It showed the ratio of siNogo-B/NC from antibody microarray. EGF, VEGF-D, FGF-4 and Progranulin were increased obviously in the HUVEC-siNogo-B supernatant, whereas anti-angiogenesis cytokines MMP-19 and VEGI declined.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Microarray, Labeling

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: HUVEC-siNogo-B-CM exhibited chemoattractive, mitogenic effects on fibroblasts. (A, B) The pictures showed the scratch test of fibroblasts in 24 h. The number of migrating fibroblasts in 50% HUVEC-siNogo-B group was significantly greater than that in 50% HUVEC-NC group in 24 h the migration (C) and proliferation (D) of fibroblasts were measured by Transwell Assay and CCk-8 Assay. The proliferative activity of fibroblasts increased markedly in 50% HUVEC-siNogo-B group as compared with 50% HUVEC-NC group after 4 days of co-culturing. Data was shown as means ± SD, n=3, *P<0.05, ***P<0.001. The scale bar is 500 µm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Migration, Transwell Assay, CCK-8 Assay, Activity Assay

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: The HUVECs-siNogo-B-CM promoted migration of epidermal cells. The migration of epidermal cells was measured by scratch test (A, B) and CCk-8 Assay (C). The epidermal cells in the 50% HUVEC-siNogo-B group was significantly greater than that in the 50% HUVEC-NC group in 12 h and 36 h. The scale bar is 500 µm. The epidermal cells in 50% HUVEC-siNogo-B group increased significantly faster after 4 days co-culturing. 5% FBS was treated as positive control and HUVEC-NC, DMEM as negtive control. Data was shown as means ± SD, n=3; *P<0.05, ***P<0.001. The scale bar is 500 µm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Migration, CCK-8 Assay, Positive Control, Control

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: HUVEC-siNogo-B could survive in 48 h after injected intradermally around the excisional wound on mice. The flow cytometry and the Immunofluorescence were performed to detected the survival of HUVEC-siNogo-B (GFP+) cells. The blue spots in the top left corner represents alive cells (A). The quantity of blue spots in two injection groups was much higher compared with vehicle control group. (B) Representative immunofluorescent staining of The GFP-labeled HUVECs. GFP+DAPI+ were survival cells by initially injected.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Injection, Flow Cytometry, Immunofluorescence, Control, Staining, Labeling

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: Topical administration of HUVEC-siNogo-B accelerated wound healing on mice. A. The wound was treated with saline, HUVEC-NC and HUVEC-siNogo-B, examined at day 1, day 3, day 5, day 7, day 14 after wounding and digitally photographed. B. Quantitative analysis of wound closure demonstrating higher wound healing rate in the HUVEC-siNogo-B group. Data was shown as means ± SD, n=6; ***P<0.001.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Saline

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: The HUVEC-siNogo-B increased callipary density in wound of nude mice. (A) Representative HE staining at day 7 after wounding. (B) Representative Immunohistochemistry staining of CD31 at day 7. Statistical analysis of CD31+ newly formed vessels showed that neovascularization was more obvious in HUVEC-siNogo-B treated wounds (C). Data was shown as means ± SD, n=5; ***P<0.001. The scale bar is 100 µm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Staining, Immunohistochemistry

Journal: Development (Cambridge, England)

Article Title: Endothelial cell SMAD6 balances Alk1 function to regulate adherens junctions and hepatic vascular development.

doi: 10.1242/dev.201811

Figure Lengend Snippet: Fig. 9. SMAD6 modulates endothelial cell hypercontractility and PI3K signaling via ALK1. (A-F) HUVEC treated with non-targeting (NT), Smad6-1 and/or Alk1 siRNA were cultured in static conditions on biotinylated fibronectin and treated as indicated for 15 min (thrombin, blebbistatin) (A-C) or 21 h (740-P, wortmannin) (D-F) at 37°C. (A) VE- cadherin stain. (B) Streptavidin-488 label. (C) Quantification of streptavidin-488 area/ field of view (FOV). Data are mean±s.d. (1 data point/FOV), n=3 experimental replicates/condition. Relevant subset of significant comparisons shown. (D) VE- cadherin stain. (E) Streptavidin-488 label. (F) Quantification of streptavidin-488 area/ FOV. Data are mean±s.d. (1 data point/ FOV), n=3 experimental replicates per condition. Relevant subset of significant comparisons shown. *P<0.05; **P<0.01; ****P<0.0001; ns, not significant. One-way ANOVA with Tukey’s multiple comparisons test. Scale bars: 20 µm (A,D); 100 µm (B,E).

Article Snippet: For contractility assays, HUVEC were treated with 0.5 U/ml thrombin (Sigma-Aldrich, T7201-500UN) at 37°C for 15 min. For contractility inhibition assays, HUVEC were treated with 10 μM blebbistatin (SigmaAldrich, B0560-1MG) at 37°C for 15 min. For PI3K activation assays, HUVEC were treated with 20 μM 740Y-P (MedChemExpress, HY-P0175) at 37°C for 22 h. For PI3K inhibition assays, HUVEC were treated with 100 nM wortmannin (SelleckChem, S2758) at 37°C for 22 h. For BMP9 ligand assays, HUVECwere serum starved in Endothelial BaseMedia (Lonza CC-3162) with 0.1% fetal bovine serum (FBS) for 24 h followed by treatment with 10 ng/ml BMP9 (R&D Systems, 3209-BP-010) at 37°C for 1 h. Immediately following drug treatments, HUVEC were fixed in warm 4% PFA at RT for 4 min. Endothelial cell flow experiments Flow experiments were performed using an Ibidi pump system as previously described (Ruter et al., 2021) with adjustments as follows: HUVEC 48 h post-transfection were seeded onto fibronectin-coated Ibidi slides (μ-Slide I Luer I 0.6 mm, 80186) in flowmedium (EBM-2with 2% FBS, 1× antibioticantimycotic, and 1% nyastatin) at a density of 2×105 cells/mm2.

Techniques: Cell Culture, Staining