Journal:

Article Title: Divalent ion block of inward rectifier current in human capillary endothelial cells and effects on resting membrane potential

doi: 10.1111/j.1469-7793.1998.119bf.x

Figure Lengend Snippet: A, current traces from a HCEC recorded in response to voltage step pulses from −120 to +10 mV in 20 mV intervals at a holding potential of −60 mV. B, current traces from the same voltage pulse protocol after application of 50 μm Ba2+. C, current-voltage plot of peak (•) and steady-state (▪) current in standard bath solution, and peak (○) and steady-state (□) current after application of Ba2+ for the same cell. D, current traces from a HCEC response to voltage steps of −120 mV to +10 mV from a holding potential of −60 mV with 5.4 mm K+ in the standard bath solution. E, current traces from the same voltage protocol in the absence of external K+. F, peak current-voltage plot from the currents in D and E before (•) and after (○) removing all external K+. Elimination of external K+ completely blocks the inward current.

Article Snippet: Cell culture Human capillary endothelial cells (HCEC) were obtained from Cell Systems Corporation (WA, USA), and were cultured in growth medium for human capillary endothelial cells (4M1-500, Cell Systems Corporation).

Techniques:

Journal:

Article Title: Divalent ion block of inward rectifier current in human capillary endothelial cells and effects on resting membrane potential

doi: 10.1111/j.1469-7793.1998.119bf.x

Figure Lengend Snippet: A, inward rectifier current traces of a HCEC bathed in standard bath solution. The cell was held at −60 mV and step potentials were applied from −120 to +20 mV, in 20 mV steps. B, the same cell after superfusion with standard bath solution containing 9 mm Ca2+. C, addition of 50 μm Ba2+ blocked the remaining current. D, current traces taken after washout of high external Ca2+ and Ba2+ with standard bath solution. E, current-voltage plots for control, 9 mm Ca2+, 50 μm Ba2+ and after washout.

Article Snippet: Cell culture Human capillary endothelial cells (HCEC) were obtained from Cell Systems Corporation (WA, USA), and were cultured in growth medium for human capillary endothelial cells (4M1-500, Cell Systems Corporation).

Techniques: Control

Journal:

Article Title: Divalent ion block of inward rectifier current in human capillary endothelial cells and effects on resting membrane potential

doi: 10.1111/j.1469-7793.1998.119bf.x

Figure Lengend Snippet: A, effect of [Ca2+]o from 0.5 to 30 mm on the mean percentage block of peak inward rectifier current at −120 mV (n = 5). Inhibition of the current at different concentrations of external Ca2+ is well fitted by a logistic equation with an EC50 value of 5.4 ± 0.6 mm. B, representative current traces of a HCEC at −120 mV with [Ca2+]o from 0 to 9 mm. C, shifts in zero current level of a HCEC exposed to divalent cations. The cell was held at −60 mV and voltage ramps were applied from −120 to +20 mV. The zero current level occurred at −62 mV with 0.5 mm Ca2+, −47 mV with 7 mm Ca2+, and −18 mV with 50 μm Ba2+ in the bath solution.

Article Snippet: Cell culture Human capillary endothelial cells (HCEC) were obtained from Cell Systems Corporation (WA, USA), and were cultured in growth medium for human capillary endothelial cells (4M1-500, Cell Systems Corporation).

Techniques: Blocking Assay, Inhibition

Journal:

Article Title: Divalent ion block of inward rectifier current in human capillary endothelial cells and effects on resting membrane potential

doi: 10.1111/j.1469-7793.1998.119bf.x

Figure Lengend Snippet: A, current traces from a HCEC in Mn2+-free standard bath solution. The cell was held at −60 mV and pulsed from −120 mV to +20 mV in 20 mV steps. B, current traces after switching to a bath solution containing 5 mm Mn2+. C, the current-voltage plot of the same cell in standard bath solution and in bath solution containing 5 mm Mn2+. D, current traces (same voltage protocol as A) of another HCEC in standard bath solution without Mg2+. E, current traces after application of 5 mm Mg2+ in the standard bath solution to the same HCEC. F, plot of the current-voltage relationship of the HCEC before and after application of Mg2+.

Article Snippet: Cell culture Human capillary endothelial cells (HCEC) were obtained from Cell Systems Corporation (WA, USA), and were cultured in growth medium for human capillary endothelial cells (4M1-500, Cell Systems Corporation).

Techniques:

Journal:

Article Title: Divalent ion block of inward rectifier current in human capillary endothelial cells and effects on resting membrane potential

doi: 10.1111/j.1469-7793.1998.119bf.x

Figure Lengend Snippet: A, IK(IR) traces during strong hyperpolarizing pulses (−180 to +20 mV) from a holding potential of −60 mV from a HCEC perfused with standard external solution. B, current traces from the same cell in the presence of 5 mm Sr2+. Both the transient and steady-state components of the inward current were blocked. C, current-voltage plot for the peak current (▪) and the current at the end of the pulse (•) in standard bath solution, and the peak current (□) and the steady-state current (○) in 5 mm Sr2+.

Article Snippet: Cell culture Human capillary endothelial cells (HCEC) were obtained from Cell Systems Corporation (WA, USA), and were cultured in growth medium for human capillary endothelial cells (4M1-500, Cell Systems Corporation).

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Comparative Evaluation of Cigarette Smoke and a Heated Tobacco Product on Corneal Oxidative Stress in an Air/Liquid Interface Model

doi: 10.1167/iovs.66.4.4

Figure Lengend Snippet: The 1R6F exposure impairs epithelial cell wound closure ability. (A) Wound healing assay on HCEC following ALI exposure to air, HTP and 1R6F. Images are representative of four time points 0, 24, 48, and 72 hours. (B) Wound healing assay on SIRC after ALI exposure to air, HTP and 1R6F. Images are representative of three time points 0, 24, and 48 hours. (C) Quantification of wound closure percentage on HCEC and (D) SIRC exposed to air, HTP and 1R6F. (** P ≤ 0.01; **** P ≤ 0.0001).

Article Snippet: Immortalized human corneal epithelial cells (HCEC) (P10863-IM; Innoprot, Bizkaia, Spain) were cultured with endothelial cell medium (P60104; Innoprot) according to manufacturer's instruction.

Techniques: Wound Healing Assay

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F expression is regulated by hypoxia in cardiac endothelial cells. (A) The expression of Sema3F RNA was increased in endothelial cells (HCECs) under hypoxic conditions, with the highest levels being observed after 4 h. HCECs were exposed to hypoxia (1 % O 2 ) for the indicated times, after which the cells were harvested, RNA was isolated and reverse transcribed. cDNA was then used for the quantification of Sema3F expression by qRT-PCR. The data presented here is derived from four independent experiments (biological replicates, n = 4), and the statistical analysis employed was the Man-Whitney test. A p-value <0.05 is considered to be significant and marked with an asterisk (*). (B) Sema3F protein was significantly increased in response to 24 h of hypoxia in comparison to normoxia. For Western blot analysis HCECs were lysed and Sema3F protein levels were detected by an anti-Sema3F antibody. GAPDH was used as a loading control. Representative blots from four independent experiments are presented (biological replicates, n = 4). The results were analyzed using the Mann-Whitney test (*p < 0.05).

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Expressing, Isolation, Reverse Transcription, Quantitative RT-PCR, Derivative Assay, Comparison, Western Blot, Control, MANN-WHITNEY

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F is an inhibitor of angiogenesis in Matrigel® sprouting assays. (A-C) In cardiac endothelial cells (HCECs) Sema3F abolished the pro-angiogenic effect of VEGF. HCECs were cultured with the indicated amounts of Sema3F or VEGF alone or in combination over 16 to 18 h. HCECs were then assayed on Matrigel® for further 2.5 h. (A) Representative micrographs of stimulated HCECs are shown. (B&C) Sema3F prevented VEGF-induced cumulative sprout length (B) and branching of HCECs (C). Data from 4 independent experiments were shown (n, number of biological replicates). The results were analyzed using the one-way ANOVA and Bonferroni's multiple comparisons test: * p < 0.05. Figs. D—F show the effect of Sema3F knockdown on angiogenesis of HCECs. The siRNA-based Sema3F knockdown resulted in a potentiated pro-angiogenic effect of VEGF. HCECs transfected with Sema3F-specific siRNA (siSema3F) or control siRNA (ctrl-siRNA) were treated with 100 ng/ml VEGF or EBM with 2 % FCS for 18 h before being assayed on Matrigel®. (D) Representative micrographs are shown. Sema3F knockdown led to a significant increase in cumulative sprout length (E) and number of branch points (F). (E&F) Data from 4 independent experiments were shown (n, number of biological replicates). All box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using one-way ANOVA followed by Bonferroni's multiple comparison test: * p < 0.05.

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Cell Culture, Knockdown, Transfection, Control, Comparison

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F inhibits angiogenesis in spheroid sprouting and aortic ring assay. (A&B) For the Spheroid sprouting assay, HCECs were formed to spheroids and were then stimulated for 24 h with the indicated amounts of Sema3F, VEGF or both. (A) Representative micrographs are shown. (B) Sema3F caused a significant decrease in sprout length and reduced the proangiogenic effect of VEGF. Data from 4 independent experiments are shown. Ten spheroids per well per condition were measured and the average cumulative sprout length was estimated (n, number of biological replicates). Analysis with one-way ANOVA followed by Bonferroni's multiple comparisons test. *p < 0.05. (C&D) In the aortic ring assay, aortic rings were stimulated with the indicated amounts of Sema3F, VEGF or both on days 3, 5 and 7. The number of capillaries was counted on day 9. (C) Compared to controls, VEGF alone increased sprouting, while co-treatment of aortic rings with Sema3F prevented VEGF-induced sprouting ex vivo. Data from 3 independent experiments, total of 10–12 aortic rings per group (n, number of biological replicates). All box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using one-way ANOVA followed by Bonferroni's multiple comparisons test. * p < 0.05.

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Aortic Ring Assay, Ex Vivo