Journal: Nutrients

Article Title: JNK/p66Shc/ITCH Signaling Pathway Mediates Angiotensin II-induced Ferritin Degradation and Labile Iron Pool Increase

doi: 10.3390/nu12030668

Figure Lengend Snippet: Angiotensin II (Ang II) induces ferritin degradation. Immunoblotting analysis of ferritin H and L levels in lysates of ( a ) human umbilical vein endothelial cells (HUVEC), ( b ) HT22, and ( c ) CPAE cells treated with 0.4 μmol/L Ang II for ( a ) 4 h, ( b ) 1 h, and ( c ) 1 h. After probing with specific antibodies, the blots were stripped and reprobed with an anti-β-actin antibody to normalize for differences in protein loading. The results are representative of three independent experiments. ( d ) Immunoblotting analysis of ferritin L levels in a lysate of CPAE cells treated with 0.4 μmol/l Ang II for the indicated time periods. After probing with specific antibodies, the blots were stripped and reprobed with an anti-β-actin antibody to normalize for differences in protein loading. Data in the graph on the right are the mean ± SEM from three independent experiments, with values recalculated relative to the control; “a” indicates statistical significance at p < 0.05 (t-test).

Article Snippet: Human umbilical vein endothelial cells (HUVEC, 200-05N) were purchased from the European Collection of Authenticated Cell Cultures (Porton Down, UK) and cultured in endothelial cell growth medium (211–500).

Techniques: Western Blot, Control

Journal: The Journal of Biological Chemistry

Article Title: AMP-dependent Kinase Inhibits Oxidative Stress-induced Caveolin-1 Phosphorylation and Endocytosis by Suppressing the Dissociation between c-Abl and Prdx1 Proteins in Endothelial Cells *

doi: 10.1074/jbc.M113.460832

Figure Lengend Snippet: AICAR suppresses phosphorylation of caveolin-1 and c-Abl, and albumin endocytosis under oxidative stress. A, cells were cultured in AICAR (2 mm) containing medium for 2 h and then stimulated with each different concentration (0, 0.5, 1.0, 2.0 mm) of H2O2 for 30 min. The amounts of p-caveolin-1 and p-c-Abl in HUVEC were then examined by Western blotting. B, densitometry of p-caveolin-1 in A is shown. C, densitometry of p-c-Abl in A. D, shown is an albumin endocytosis assay. a, control (untreated cells). b, H2O2 (2 mm) stimulation for 30 min. c, pretreated with AICAR (2 mm) for 2 h. d, pretreated with AICAR for 2 h followed by H2O2 (2 mm) stimulation for 30 min. BSA was conjugated with Alexa 555 (red), p-caveolin (green), VE-cadherin (blue). Scale bar = 50 μm. A, representative blots are shown. *, p < 0.01.

Article Snippet: Cell Culture HUVECs (Lonza, Walkersville, MD) were cultured in endothelial growth medium (Lonza, Walkersville, MD).

Techniques: Phospho-proteomics, Cell Culture, Concentration Assay, Western Blot, Endocytosis Assay, Control

Journal: Cell Proliferation

Article Title: Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity

doi: 10.1046/j.1365-2184.2001.00205.x

Figure Lengend Snippet: Correlation between the number of HUVEC (a, b) and MIEC (c, d) with the optical density of the WST‐1 reaction product measured after 24 h in basal medium with FCS (a, c) and complete medium with FCS (b, d).

Article Snippet: Cell culture HUVEC and MIEC were purchased from BioWhittaker‐Clonetics (Verviers, Belgium) and cultured in BioWhittaker‐Clonetics complete medium® containing 5% (v/v) dialysed fetal bovine serum, bovine brain extract (18 μg/ml), rhEGF (10 ng/ml), hydrocortisone (1.0 μg/ml), gentamycin (50 mg/ml) and amphotericin B (50 ng/ml).

Techniques:

Journal: Cell Proliferation

Article Title: Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity

doi: 10.1046/j.1365-2184.2001.00205.x

Figure Lengend Snippet: Influence of seeding density and FCS on the growth characteristics of HUVEC and MIEC in the absence of cytokines. (a) 3000 and (b) 6000 cells/well were seeded in the presence or absence of FCS and the optical density of the WST‐1 reaction product was measured after 24 h (□), 48 h (▒) and 72 h (▪). In the presence of serum all control cultures exhibited a significant increase in cell density, whereas in the absence of serum only HUVEC proliferated. n.d. not determined.

Article Snippet: Cell culture HUVEC and MIEC were purchased from BioWhittaker‐Clonetics (Verviers, Belgium) and cultured in BioWhittaker‐Clonetics complete medium® containing 5% (v/v) dialysed fetal bovine serum, bovine brain extract (18 μg/ml), rhEGF (10 ng/ml), hydrocortisone (1.0 μg/ml), gentamycin (50 mg/ml) and amphotericin B (50 ng/ml).

Techniques: Control

Journal: Cell Proliferation

Article Title: Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity

doi: 10.1046/j.1365-2184.2001.00205.x

Figure Lengend Snippet: Dose‐dependent effects of cytokines on HUVEC (a) and MIEC (b) expressed as percentage increase over untreated controls. Six‐thousand cells/well were incubated for 72 h in basal medium with 2.5% FCS containing various amounts (0.1–100 ng/ml) of VEGF 121, VEGF 165, PlGF‐1, PlGF‐2 and FGF‐2. (c) Comparison of the mitogenic effects of the cytokines at a concentration of 100 ng/ml between HUVEC (▪) and MIEC (▒). At this concentration VEGF 121, VEGF 165 and FGF‐2 are potent cell mitogens, PlGF‐1 and PlGF‐2 did not produce a significant effect under these conditions, except for a weak proliferative effect of PlGF‐1 on MIEC. *P < 0.05, #P < 0.01, §P < 0.001 vs. untreated control.

Article Snippet: Cell culture HUVEC and MIEC were purchased from BioWhittaker‐Clonetics (Verviers, Belgium) and cultured in BioWhittaker‐Clonetics complete medium® containing 5% (v/v) dialysed fetal bovine serum, bovine brain extract (18 μg/ml), rhEGF (10 ng/ml), hydrocortisone (1.0 μg/ml), gentamycin (50 mg/ml) and amphotericin B (50 ng/ml).

Techniques: Incubation, Comparison, Concentration Assay, Control

Journal: Cell Proliferation

Article Title: Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity

doi: 10.1046/j.1365-2184.2001.00205.x

Figure Lengend Snippet: Kinetics of the cytokine effects on HUVEC (▪) and MIEC (▒) expressed as percentage increase over untreated controls. Six‐thousand cells/well were incubated for 24 h (a) and 48 h (b) in serum‐free basal medium in the presence or absence (control) of 100 ng/ml of the respective growth factor. In general, the response of MIEC increases, whereas that of HUVEC decreases from 24 to 48 h. *P < 0.05, #P < 0.01, §P < 0.001 vs. untreated control.

Article Snippet: Cell culture HUVEC and MIEC were purchased from BioWhittaker‐Clonetics (Verviers, Belgium) and cultured in BioWhittaker‐Clonetics complete medium® containing 5% (v/v) dialysed fetal bovine serum, bovine brain extract (18 μg/ml), rhEGF (10 ng/ml), hydrocortisone (1.0 μg/ml), gentamycin (50 mg/ml) and amphotericin B (50 ng/ml).

Techniques: Incubation, Control

Journal: Cell Proliferation

Article Title: Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity

doi: 10.1046/j.1365-2184.2001.00205.x

Figure Lengend Snippet: Serum‐dependent effects of the cytokines on HUVEC (▪) and MIEC (▒) expressed as percentage increase over untreated controls. Three thousand cells/well were incubated for 24 h in serum‐free basal medium (a) or in medium with FCS (b), in the presence or absence (control) of 100 ng/ml of the respective growth factor. The proliferative response of MIEC to growth factors is more dependent on serum presence than that of HUVEC. Note that at this seeding density PlGF‐1 and PlGF‐2 elicited a mitogenic effect also in HUVEC *P < 0.05, #P < 0.01, §P < 0.001 vs. untreated control.

Article Snippet: Cell culture HUVEC and MIEC were purchased from BioWhittaker‐Clonetics (Verviers, Belgium) and cultured in BioWhittaker‐Clonetics complete medium® containing 5% (v/v) dialysed fetal bovine serum, bovine brain extract (18 μg/ml), rhEGF (10 ng/ml), hydrocortisone (1.0 μg/ml), gentamycin (50 mg/ml) and amphotericin B (50 ng/ml).

Techniques: Incubation, Control