Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Endothelial Cell Growth Medium MV 2 , Promo Cell , C-22121.

Techniques: Recombinant, Protease Inhibitor, Staining, Western Blot, Membrane, Protein Extraction, Plasmid Preparation, Software, Imaging, Microscopy

Journal: bioRxiv

Article Title: Spatial expression of an mRNA encoding Tie2-agonist in the capillary endothelium of the lung prevents pulmonary vascular leakage

doi: 10.1101/2022.10.12.511878

Figure Lengend Snippet: (A) Effect of COMP-Ang1 on F-actin remodeling and VE-Cadherin distribution in isolated human pulmonary microvascular endothelial cell. HPMECs grown on glass coverslips were transfected for 24h with control mRNA LUC or with mRNA-76 encoding COMP-Ang1 with or without temporal addition of 10 ng/ml TNFα, followed by immunofluorescence staining with VE-Cadherin-specific antibody and with Alexa Fluor 594 phalloidin to detect actin filaments. Arrows indicate disrupted VE-Cadherin complexes. B) Transcellular electrical resistance (TER) measurements of HPMEC monolayers. HPMEC were incubated for 30 min (top) or 60 min (bottom) with supernatant from mRNA-76 or with mRNA LUC (Luciferase) and the treated with supernatants from Hela cells transfected for 24 h with control mRNA LUC or mRNA-76 encoding COMP-Ang1 and stimulated with indicated amount of pneumolysin (PLY; 0.25, 0.5, 0.75, 1.0 μg/ml). PLY stimulation decreased TER of HPMEC monolayers, displaying loss of endothelial integrity when incubated with control Luciferase supernatant (solid curve). Preincubation with COMP-Ang1 containing supernatant attenuated the PLY-induced TER decrease (purple lines dashed versus solid).

Article Snippet: Media containing formulations was removed and replaced with 10 % MV2 growth medium for 16 h. For TNFα treatment, 10 % MV2 growth medium with TNFα (R&D Systems, cat. #210-TA) or vehicle was spiked into each well for a final concentration of 10 ng/ml for a further 6 h. Cells were fixed in 4 % PFA (Thermo Scientific, cat. #15670799) for 10 min at room temperature (RT).

Techniques: Isolation, Transfection, Control, Immunofluorescence, Staining, Incubation, Luciferase