Journal: PLoS ONE

Article Title: E-Selectin Mediated Adhesion and Migration of Endothelial Colony Forming Cells Is Enhanced by SDF-1α/CXCR4

doi: 10.1371/journal.pone.0060890

Figure Lengend Snippet: A) LPS-treated HUVEC cells demonstrate significant cellular injury and increased lactate dehydrogenase (LDH) after 3h (** p<0.01). B) Gene expression of E-selectin, VCAM-1, ICAM-1 and SDF-1α induced in HUVECs after endotoxic injury was determined by RT-PCR relative to GAPDH expression (* denotes p<0.05 and ** p<0.01 in comparison with baseline). C) E-selectin levels after 12 hours in conditioned media from LPS-treated HUVEC cells (H LPS ) compared with ECFCs exposed to conditioned media from control HUVEC cells (H) or LPS-treated HUVECs cultured in the presence of anti-CD62E which inhibits soluble E-selectin (H+anti-CD62E) LPS , as measured using ELISA.(** p<0.01)

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) were obtained from the supplier (Lonza) and cultured according to instructions.

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Expressing, Comparison, Control, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: E-Selectin Mediated Adhesion and Migration of Endothelial Colony Forming Cells Is Enhanced by SDF-1α/CXCR4

doi: 10.1371/journal.pone.0060890

Figure Lengend Snippet: A) ECFC adhesion to fibronectin following exposure to conditioned media from control HUVEC cells (H), from injured HUVEC cells (H LPS ), and from LPS-treated HUVECs with the addition of anti-CD62E (H+anti-CD62E) LPS and shown as cells per high powered field (hpf) (* p<0.05). B) Adhesion of ECFCs to fibronectin following 2 hour incubation with E-selectin at 2 µg/ml (E2) or 4 µg/ml (E4) compared to controls (0 µg/ml, E0), in the presence of anti-CD44 and E-selectin (4 µg/ml) and VEGF-A 100 ng/ml (VEGFA). (* p<0.05). C) Migration of ECFCs towards E-selectin in the lower chamber at 2 µg/ml (E2) or 4 µg/ml (E4) compared with controls containing no E-selectin in the lower chamber (E0) or when ECFCs were incubated in the presence of anti-CD44 with E-selectin 4 µg/ml in the lower chamber (A+E4), and to SDF-1a (100 ng/ml) in the lower chamber (SDF). (* p<0.05). D) Photgraphs of the underside of porous membranes used in the migration assays described in part C.

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) were obtained from the supplier (Lonza) and cultured according to instructions.

Techniques: Control, Incubation, Migration

Journal: PLoS ONE

Article Title: E-Selectin Mediated Adhesion and Migration of Endothelial Colony Forming Cells Is Enhanced by SDF-1α/CXCR4

doi: 10.1371/journal.pone.0060890

Figure Lengend Snippet: A) SDF-1α mRNA expression was assessed after endotoxic injury in HUVECs with an early decrease in expression that was evident at 3 hours and persisted up to 12 hours. (* p>0.05). B) Following LPS treatment of MRC5 cells, mRNA expression of SDF-1a is increased after 6h (* p<0.05). Gene expression is reported relative to GAPDH.

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) were obtained from the supplier (Lonza) and cultured according to instructions.

Techniques: Expressing, Gene Expression

Journal: PLoS ONE

Article Title: E-Selectin Mediated Adhesion and Migration of Endothelial Colony Forming Cells Is Enhanced by SDF-1α/CXCR4

doi: 10.1371/journal.pone.0060890

Figure Lengend Snippet: A) E-selectin expression in HUVEC cells following endotoxic (diamonds), hypoxic (squares) and radiation injury (triangles). ** p<0.01 for endotoxic injury at 3h compared to 0h. B) SDF-1α expression in HUVEC cells following endotoxic (diamonds), hypoxic (squares) or radiation injury (triangles). ** p<0.01 for hypoxic injury at 6h compared to 0h. C) SDF-1α expression following endotoxic injury to MRC5 cells (triangles) and HUVEC cells (squares), ** p<0.01 when comparing expression levels at 6h. D) SDF-1α expression following hypoxic injury to MRC5 cells (triangles) and HUVEC cells (squares), ** p<0.01 comparing expression levels at 6h; * p<0.05 comparing expression levels at 6 hours in MRC5 cells to 0 hours.

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) were obtained from the supplier (Lonza) and cultured according to instructions.

Techniques: Expressing

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Lixisenatide protects oxygen-glucose deprivation/reperfusion (OGD/R)-induced reduction of cell viability and release of LDH in HUVECs. Cells were treated with lixisenatide (5, 10, 20 nM) for 6 h, followed by exposure to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). (A) Cell viability of HUVECs; (B) release of lactate dehydrogenase (LDH); (C) cell morphology of HUVECs (****, P < 0.0001 vs. control group; ## , ### , #### , P < 0.01, 0.001, 0.0001 vs. OGD/R treatment group, n = 5).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: Control

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Lixisenatide reduced OGD/R-induced generation of ROS in HUVECs. Cells were treated with lixisenatide (10, 20 nM) for 6 h, followed by exposure to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). Generation of ROS was measured by dihydroethidium (DHE) staining (****, P < 0.0001 vs. control group; ### , #### , P < 0.001, 0.0001 vs. OGD/R treatment group, n = 5).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: Staining, Control

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Lixisenatide activated the expression of Nrf2 and HO-1 under OGD/R in HUVECs. Cells were treated with lixisenatide (10, 20 nM) for 6 h, followed by exposure to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). Expression of Nrf2 and HO-1 was measured by western blot analysis (**, ***, ****, P < 0.01, 0.001, 0.0001 vs. control group, n = 4).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: Expressing, Western Blot, Control

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Lixisenatide prevented OGD/R-induced reduced angiogenesis in HUVECs. Cells were treated with lixisenatide (20 nM) for 6 h, followed by exposure to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). Microtubule formation was quantified by Matrigel assay (****, P < 0.0001 vs. control group; #### , P < 0.0001 vs. OGD/R treatment group, n = 5).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: Matrigel Assay, Control

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Lixisenatide prevented OGD/R-induced dephosphorylation of PI3K and Akt in HUVECs. Cells were treated with lixisenatide (20 nM) for 6 h, followed by exposure to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). Phosphorylated and total levels of PI3K and Akt were measured by western blot analysis (****, P < 0.0001 vs. control group; #### , P < 0.0001 vs. OGD/R treatment group, n = 4).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: De-Phosphorylation Assay, Western Blot, Control

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Lixisenatide prevented OGD/R-induced dephosphorylation of eNOS and production of nitric oxide (NO) in HUVECs. Cells were treated with lixisenatide (20 nM) for 6 h, followed by exposure to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). (A) Phosphorylated and total levels of eNOS were measured by western blot analysis. (B) Intracellular nitric oxide (NO) was measured by diaminofluorescein-FM diacetate (DAF-FM DA) (****, P < 0.0001 vs. control group; ### , #### , P < 0.01, 0.001, 0.0001 vs. OGD/R treatment group, n = 5).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: De-Phosphorylation Assay, Western Blot, Control

Journal: RSC Advances

Article Title: The protective effects of GLP-1 receptor agonist lixisenatide on oxygen-glucose deprivation/reperfusion (OGD/R)-induced deregulation of endothelial tube formation

doi: 10.1039/c9ra09959j

Figure Lengend Snippet: Blockage of the PI3K/Akt signaling pathway using LY294002 abolished the protective effects of lixisenatide on promoting angiogenesis in HUVECs. Cells were treated with lixisenatide (20 nM) in the presence or absence of LY294002 (20 μM) for 6 h, followed by exposed to oxygen-glucose deprivation (6 h)/reperfusion (24 h) (OGD/R). (A) Molecular structure of LY294002; (B) microtubule formation was quantified by Matrigel assay; (C) production of ROS (****, P < 0.0001, n = 5).

Article Snippet: Human umbilical vascular endothelial cells (HUVECs), culture medium (EGM-2), growth factor BulletKits, and 0.25% trypsin–EDTA were purchased from Lonza (USA).

Techniques: Matrigel Assay