Journal: PLoS ONE

Article Title: Twist1 Controls Lung Vascular Permeability and Endotoxin-Induced Pulmonary Edema by Altering Tie2 Expression

doi: 10.1371/journal.pone.0073407

Figure Lengend Snippet: A ) Immunofluorescence micrographs showing cell-cell junction structure by VE-cadherin staining in L-HMVE cells treated with Twist1 siRNA #1 or #2 (bar, 50 µm). DAPI staining shows the nuclei of cells. Arrows show the region where cell-cell junctions are disrupted. As a control, cells were treated with siRNA duplex with an irrelevant sequence. B ) Graph showing the quantitation of the total discontinuous area of at least 10 fields (*, p<0.01). C ) Graph showing endothelial permeability in L-HMVE cells treated with control siRNA, Twist1 siRNA #1 or #2 (*, p<0.05). Permeability (Pa) values were evaluated after 6 h and are expressed as percentage of control cells. D ) Immunoblots showing tyrosine phosphorylated Tie2 detected by phosphotyrosine antibody (4G10) and total Tie2 immunoprecipitated with Tie2 antibody from total cell lysates in Twist1 knockdown (siRNA #1 treated) L-HMVE cells. E ) Levels of active RhoA in Twist1 knockdown (siRNA #1 treated) L-HMVE cells. Quantitative results (ratio of active RhoA to total RhoA) were normalized to control siRNA treated cells (*, p<0.01). F ) Graph showing endothelial permeability in HUVE cells treated with Twist1 siRNA #1, Tie2 DNA, or in combination (*, p<0.05). As a control, cells were treated with siRNA duplex with an irrelevant sequence and/or control DNA (vector only). Immunoblots showing Twist1, Tie2, and GAPDH protein levels in HUVE cells treated with Twist1 siRNA #1, Tie2 DNA, or in combination. All error bars are s.e.m.

Article Snippet: L-HMVE cells and HUVE cells (Lonza, Walkersville, MD) were cultured as described before [ , ].

Techniques: Immunofluorescence, Staining, Sequencing, Quantitation Assay, Permeability, Western Blot, Immunoprecipitation, Plasmid Preparation

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: Oxidized low-density lipoprotein (oxLDL) induces proliferation in human umbilical vein endothelial cells (HUVECs) by CCK-8 assay. OxLDL at concentrations of 10 μg/ml, 25 μg/ml, 50 μg/ml, 75 μg/ml, 100 μg/ml were applied to HUVECs for 3 h (A), 6 h (B), 12 h (C), 24 h (D), 48 h (E) with basal conditions, and then the optical density (OD) at 450 nm was measured. Each data point represents mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01 compared with control. #P<0.05, ##P<0.01 compared with normal.

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: CCK-8 Assay, Standard Deviation, Control

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: The effects of native LDL and oxLDL on HUVEC. Both native LDL and oxLDL at concentrations of 10 μg/ml, 25 μg/ml, 50 μg/ml, 75 μg/ml, 100 μg/ml for 24 h were applied to HUVECs. The impact of native LDL or oxLDL on the morphological changes in HUVECs using phase-contrast microscope (×100) (A, B, respectively) and on the proliferation using CCK-8 (C). Each data point represents mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01 compared with control.

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: Microscopy, CCK-8 Assay, Standard Deviation, Control

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: Berberine inhibits proliferation of HUVEC induced by oxLDL. Pretreated with berberine at concentrations of 5 μg/ml, 10 μg/ml, 25 μg/ml, 50 μg/ml, attenuated the oxLDL-induced proliferation of HUVECs in a dose-dependent manner using CCK-8 assay (A). Berberine at concentration of 25 μg/ml significantly inhibited 50 μg/ml oxLDL-induced proliferation of HUVECs using EdU proliferation assay using a confocal laser scanning microscope (×100) (B) and the ratio of EdU-positive cells to total cells was shown (C). Each data point represents mean ± standard deviation of three independent experiments. **P<0.01 compared with control. #P<0.05, ##P<0.01 compared with oxLDL 50 μg/ml. $P<0.05 compared with Control.

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: CCK-8 Assay, Concentration Assay, Proliferation Assay, Laser-Scanning Microscopy, Standard Deviation, Control

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: Berberine significantly decreased PCNA, NF-кB, LOX-1 and PI3K mRNA expression induced by oxLDL in HUVECs in a dose-dependent manner. Real-time PCR assay for PCNA (A), NF-кB (B), LOX-1 (C) and PI3K (D), respectively. Each data point represents mean ± standard deviation of three independent experiments. **P<0.01 compared with control. ##P<0.01 compared with oxLDL 50 μg/ml.

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Control

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: OxLDL induced the proliferation through up-regulating PCNA, LOX-1 and activating NF-кB, phospho-Akt, phospho-ERK, phospho-p38 signal pathways in a dose-dependent manner. Western blot to PCNA, NF-кB, LOX-1 (A, E, F, G), and phospho-Akt (B, H), phospho-ERK (C, I), phospho-p38 (D, J) in HUVECs. Each data point represents mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01 compared with control.

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: Western Blot, Standard Deviation, Control

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: Berberine inhibited oxLDL-induced HUVECs proliferation by down-regulating the PCNA, LOX-1 and inactivating NF-кB, phospho-Akt, phospho-ERK, phospho-p38 signal pathways. Western blot to PCNA, NF-кB, LOX-1 (A, E, F, G), and phospho-Akt (B, H), phospho-ERK (C, I), phospho-p38 (D, J) in HUVECs. Each data point represents mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01 compared with control. #P<0.05, ##P<0.01 compared with oxLDL 50 μg/ml.

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: Western Blot, Standard Deviation, Control

Journal: American Journal of Translational Research

Article Title: Impacts of berberine on oxidized LDL-induced proliferation of human umbilical vein endothelial cells

doi:

Figure Lengend Snippet: Berberine inhibited oxLDL-induced HUVECs proliferation through PI3k/Akt, ERK and p38MAPK signal pathways. HUVECs were pretreated with or without LY294002 (25 μM), PD98059 (10 μM) and SB202190 (10 μM) for 1 h followed by berberine (25 μg/ml) incubation with oxLDL (50 μg/ml) induction for 24 h. A-F: Western blot analysis showed a clear increase of Akt, ERK and p38 phosphorylation induced by LY294002, PD98059 and SB202190. G: CCK-8 assay showd that LY294002, PD98059 and SB202190 abrogated the inhibition of berberine in oxLDL-induced HUVECs proliferation. Each data point represents mean ± standard deviation of three independent experiments. **P<0.01 compared with control. #P<0.05, ##P<0.01 compared with oxLDL 50 μg/ml. $P<0.05, $$P<0.01, compared with berberine (25 μg/ml).

Article Snippet: Culture of HUVEC HUVECs were purchased from Sciencell and grown in M199 with 10% (v/v) FBS, 10 ng/ml ECGF and 100 U/ml penicillin-streptomycin (complete medium) at 37°C in a humidified atmosphere containing 5% CO 2 and 95% air.

Techniques: Incubation, Western Blot, Phospho-proteomics, CCK-8 Assay, Inhibition, Standard Deviation, Control

Journal: Tissue Engineering and Regenerative Medicine

Article Title: A Novel Hypothesis and Characterization to Isolate Microvascular Endothelial Cells Simultaneously with Adipose-Derived Stem Cells from the Human Adipose-Derived Stromal Vascular Fraction

doi: 10.1007/s13770-021-00332-5

Figure Lengend Snippet: Comparative analysis of properties according to the HMVEC-A isolation method. A Morphology and tube formation of HMVEC-A isolated by the vascular fraction (VF) isolation, magnetic-activated cell sorting (MACS) isolation method, and fluorescence-activated cell sorting (FACS) isolation methods were compared with Lonza HUVEC and HMVEC by phase-contrast microscopy; magnification, 40 × . B The VF isolation method produced a significantly higher yield than the FACS isolation method and MACS isolation method (*p < 0.05, **p < 0.01, n = 3). The doubling time was in the order of VF isolation method, MACS isolation method and FACS isolation method, and VF showed the fastest doubling time, but there was no statistically significant difference. The error bars indicate the standard errors of the mean of triplicate measurements. C FACS verification of EC-specific marker expression according to each isolation method. Positivity for the endothelial-specific cell surface antigens CD31 (PECAM1) and CD146 (MCAM) is indicated. Cells were analyzed at passage 2 (n = 3)

Article Snippet: Lonza HUVEC and HMVEC were used as controls.

Techniques: Isolation, FACS, Fluorescence, Microscopy, Produced, Marker, Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: A Novel Hypothesis and Characterization to Isolate Microvascular Endothelial Cells Simultaneously with Adipose-Derived Stem Cells from the Human Adipose-Derived Stromal Vascular Fraction

doi: 10.1007/s13770-021-00332-5

Figure Lengend Snippet: Comparative analysis of properties according to the HMVEC-A isolation method. A Morphology and tube formation of HMVEC-A isolated by the vascular fraction (VF) isolation, magnetic-activated cell sorting (MACS) isolation method, and fluorescence-activated cell sorting (FACS) isolation methods were compared with Lonza HUVEC and HMVEC by phase-contrast microscopy; magnification, 40 × . B The VF isolation method produced a significantly higher yield than the FACS isolation method and MACS isolation method (*p < 0.05, **p < 0.01, n = 3). The doubling time was in the order of VF isolation method, MACS isolation method and FACS isolation method, and VF showed the fastest doubling time, but there was no statistically significant difference. The error bars indicate the standard errors of the mean of triplicate measurements. C FACS verification of EC-specific marker expression according to each isolation method. Positivity for the endothelial-specific cell surface antigens CD31 (PECAM1) and CD146 (MCAM) is indicated. Cells were analyzed at passage 2 (n = 3)

Article Snippet: When the most important characteristic of HMVEC-A—i.e., their vascularity—was evaluated, we found that the level of HMVEC-A tube formation was similar to that of Lonza HUVEC and HMVEC.

Techniques: Isolation, FACS, Fluorescence, Microscopy, Produced, Marker, Expressing

Journal: Theranostics

Article Title: Nerve modulation therapy in gouty arthritis: targeting increased sFRP2 expression in dorsal root ganglion regulates macrophage polarization and alleviates endothelial damage

doi: 10.7150/thno.33908

Figure Lengend Snippet: Neural modulation of macrophages induces HUVEC apoptosis. (A), HUVECs cultured with cocultured CM from macrophages and DRG/transduced DRG neurons in the presence of MSU. Apoptosis was assessed by the TUNEL assay in HUVECs. Scale bar, 100 μm. a. The percentage of TUNEL-positive nuclei was quantified. (B), HUVECs were stained with FITC Annexin V and propidium iodide (PI) for flow cytometry analysis after being cultured with cocultured CM from macrophages and DRG/transduced DRG neurons in the presence of MSU. b. The apoptosis rate was quantified. (C), qRT-PCR was performed to assess the expression of E-selectin, ICAM-1, VCAM-1, and MCP-1 in HUVECs after being cultured with cocultured CM from macrophages and DRG/transduced DRG neurons in the presence of MSU. In vitro experiments were conducted in biological triplicate. Values are the means ± SD (*P < 0.05 compared with control. **P<0.01 compared with control. All statistical significance was determined using one-way ANOVA and Tukey's post comparison test.

Article Snippet: The assay was conducted according to manufacturer's protocol using Bio-Plex Pro Mouse Cytokine Grp (#M60009RDPD) with Luminex 200 system (Austin, TX, USA) in Wayen Biotechnologies Shanghai, Inc. HUVEC (HUVEC-20001) were purchased from Cyagen Biosciences and cultured in MEM-α (Invitrogen, 11090081) supplemented with 10% FBS.

Techniques: Cell Culture, TUNEL Assay, Staining, Flow Cytometry, Quantitative RT-PCR, Expressing, In Vitro, Control, Comparison

Journal: Tissue Engineering and Regenerative Medicine

Article Title: A Novel Hypothesis and Characterization to Isolate Microvascular Endothelial Cells Simultaneously with Adipose-Derived Stem Cells from the Human Adipose-Derived Stromal Vascular Fraction

doi: 10.1007/s13770-021-00332-5

Figure Lengend Snippet: Comparative analysis of properties according to the HMVEC-A isolation method. A Morphology and tube formation of HMVEC-A isolated by the vascular fraction (VF) isolation, magnetic-activated cell sorting (MACS) isolation method, and fluorescence-activated cell sorting (FACS) isolation methods were compared with Lonza HUVEC and HMVEC by phase-contrast microscopy; magnification, 40 × . B The VF isolation method produced a significantly higher yield than the FACS isolation method and MACS isolation method (*p < 0.05, **p < 0.01, n = 3). The doubling time was in the order of VF isolation method, MACS isolation method and FACS isolation method, and VF showed the fastest doubling time, but there was no statistically significant difference. The error bars indicate the standard errors of the mean of triplicate measurements. C FACS verification of EC-specific marker expression according to each isolation method. Positivity for the endothelial-specific cell surface antigens CD31 (PECAM1) and CD146 (MCAM) is indicated. Cells were analyzed at passage 2 (n = 3)

Article Snippet: A Morphology and tube formation of HMVEC-A isolated by the vascular fraction (VF) isolation, magnetic-activated cell sorting (MACS) isolation method, and fluorescence-activated cell sorting (FACS) isolation methods were compared with Lonza HUVEC and HMVEC by phase-contrast microscopy; magnification, 40 × .

Techniques: Isolation, FACS, Fluorescence, Microscopy, Produced, Marker, Expressing