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
