Journal: PLoS ONE

Article Title: IL-4 Causes Hyperpermeability of Vascular Endothelial Cells through Wnt5A Signaling

doi: 10.1371/journal.pone.0156002

Figure Lengend Snippet: Uniform confluent monolayers of HCAEC cultured in stabilized and collagen coated ECIS 8W10E+ arrays were treated with IL-4. TEER of HCAEC monolayers was measured in Ohms continuously every 5 min at multiple frequencies ranging from 62.6 Hz to 64 kHz, normalized to its value at time zero and plotted with respect to time. Stimulations conducted in duplicate wells were grouped and averaged to plot a single curve with error bars representing mean ± SD. Figures shown depict the resistance measurements conducted at 4000 Hz and represent four independent experiments. (A) 1, non-treated Wnt5A-siRNA transfected HCAEC; 2, non-treated non-transfected HCAEC; 3, IL-4 treated Wnt5A-siRNA transfected HCAEC; 4, IL-4 treated non-transfected HCAEC. (B) 1, non-treated negative control siRNA transfected HCAEC; 2, IL-4 treated negative control siRNA transfected HCAEC; 3, IL-4 treated non-transfected HCAEC. (C) Black, non-treated; green, IL-6.

Article Snippet: Endothelial barrier function was detected in real time using the ECIS® Z-theta system (Applied Biophysics, http://www.biophysics.com/barrierfunction.php ) with associated software v.1.2.126 PC as follows: after equilibrating with EBM-2 basal medium (Clonetics, Lonza, USA) for 24 h, 8W10E+ arrays were coated with rat tail collagen type I (diluted 0.01% in sterile pyrogen-free water; BD Biosciences, USA) for 12 h. A monodisperse suspension of HCAEC in EGM-2 MV medium (Clonetics, Lonza, USA) supplemented with 5% FBS was added to the arrays at a density of 80,000–90,000 cells/well and incubated up to 24 h to form a uniform dense monolayer ( ).

Techniques: Cell Culture, Transfection, Negative Control

Journal: PLoS ONE

Article Title: IL-4 Causes Hyperpermeability of Vascular Endothelial Cells through Wnt5A Signaling

doi: 10.1371/journal.pone.0156002

Figure Lengend Snippet: Uniform confluent monolayers of HCAEC cultured in stabilized and collagen coated ECIS 8W1E cultureware were treated with IL-4 either alone or in combination with Y-27632. Three hours after treatment, wells were subjected to an elevated electric field to create a wound. Measurements were started immediately after wounding. Impedance of HCAEC monolayer was continuously measured in Ohms every 5 min at multiple frequencies ranging from 62.6 Hz to 64 kHz and plotted with respect to time. Representative figures depict the impedance of wells after wounding was applied. Figures shown depict the measurements conducted at 4000 Hz and represent four independent experiments. (A) Black, non-treated; Blue, IL-4. (B) Black, control with Y27632; Green, IL-4 + Y27632.

Article Snippet: Endothelial barrier function was detected in real time using the ECIS® Z-theta system (Applied Biophysics, http://www.biophysics.com/barrierfunction.php ) with associated software v.1.2.126 PC as follows: after equilibrating with EBM-2 basal medium (Clonetics, Lonza, USA) for 24 h, 8W10E+ arrays were coated with rat tail collagen type I (diluted 0.01% in sterile pyrogen-free water; BD Biosciences, USA) for 12 h. A monodisperse suspension of HCAEC in EGM-2 MV medium (Clonetics, Lonza, USA) supplemented with 5% FBS was added to the arrays at a density of 80,000–90,000 cells/well and incubated up to 24 h to form a uniform dense monolayer ( ).

Techniques: Cell Culture, Control