Journal: The international journal of biochemistry & cell biology

Article Title: Netrin-4 expression by human endothelial cells inhibits endothelial inflammation and senescence.

doi: 10.1016/j.biocel.2021.105960

Figure Lengend Snippet: Fig. 5. Reduction of NTN4 expression in endothelial cells diminishes endothelial barrier function and promotes an inflammatory phenotype. (A) Left graph shows transendothelial electrical resistance of scrambled shRNA (sh-Ctrl) or anti-NTN4 shRNA (sh-NTN4) transduced HUVECs seeded on electric cell–substrate impedance sensing (ECIS) electrodes (mean ± SD of technical triplicates, representative of three independent experiments). Right graph shows the relative trans-endothelial electrical resistance of sh-Ctrl or sh-NTN4 HUVEC stable monolayer, sh-Ctrl set to 1. Mean ± SEM of n = 3. (B) HUVECs transduced with sh-Ctrl or sh-NTN4 were seeded in the vertical channel of a microfluidic device with collagen in the horizontal channel. At the start of the measurement, fluorescently labeled albumin was infused in the vessel lumen and after 10 min albumin leakage outside the vessel channel was visualized and quantified. Dotted box indicates the region used in quantifica tion. Bar graph shows the permeability coefficient presented as mean ± SEM of n = 3; *p < 0.05. (C) Quantification of adhesion of primary human monocytes to sh-Ctrl or sh-NTN4 transduced HUVECs. Mean ± SEM of n = 3; *p < 0.05 (D) ICAM1 and VCAM1 mRNA expression in sh-Ctrl or sh-NTN4 transduced HUVECs. Results are presented relative to sh-Ctrl. Mean ± SEM of n = 3; *p < 0.05. (E) Immunoblots of ICAM1, VCAM1 and GAPDH (housekeeping) in sh- Ctrl and sh-NTN4 treated HUVECs. n = 4.

Article Snippet: Electric Cell-substrate Impedance Sensing (ECIS) assays were done using the ECIS Ztheta device (Applied BioPhysics) with standard 8-well arrays (Applied BioPhysics, 8W10E).

Techniques: Expressing, shRNA, Electric Cell-substrate Impedance Sensing, Transduction, Labeling, Permeability, Western Blot

Journal: Scientific Reports

Article Title: GRP78 translocation to the cell surface and O-GlcNAcylation of VE-Cadherin contribute to ER stress-mediated endothelial permeability

doi: 10.1038/s41598-019-47246-w

Figure Lengend Snippet: HRECs exposed to TNF-α, and high glucose for 24 h demonstrated decreased trans-endothelial electrical resistance. Representative ECIS tracings plotted as normalized resistance from all treatment groups ( A ). Representative confocal immunofluorescence images of the cells stained for VE-Cadherin. Boxed images shows higher magnification. White arrows indicate loss of cell-cell contacts ( B ). Data represent Mean ± SEM from 3 independent experiments performed in duplicates. **p < 0.01; *p < 0.05 compared to controls. Scale bar = 20 μm.

Article Snippet: Measurements of trans-endothelial electrical resistance (TER) were performed using an electric cell-substrate impedance sensing (ECIS) device (ECIS Zθ; Applied Biophysics, Inc., Troy, NY), as described by us previously .

Techniques: Immunofluorescence, Staining