Journal: International Journal of Molecular Sciences

Article Title: Melanoma Cells Produce Large Vesicular-Bodies That Cause Rapid Disruption of Brain Endothelial Barrier-Integrity and Disassembly of Junctional Proteins

doi: 10.3390/ijms24076082

Figure Lengend Snippet: Schematic explaining ECIS theory. ( A ) Endothelial cells grown on the gold-plated electrodes of the ECIS plate that are coated with collagen, forming a confluent monolayer. The monolayer is formed via intercellular junctions and junctions with the collagen substrate, creating the paracellular and basolateral junctions, respectively. The formation of these junctions prevents high current flow from the electrode through the cells, where the red and green arrows represent the paracellular (Rb) and basolateral (alpha) current flow, respectively. This maintained electrical resistance is shown in graph C (Black). ( B ) The endothelial monolayer following treatment with factors that disrupt the endothelial barrier, such as melanoma cells or melanoma vesicular-bodies; the junctions weaken, allowing for greater current flow through the cells. This results in a decreased electrical resistance, shown in graph ( C ) (Blue). ( C ) A model ECIS graph depicting the plateau in resistance formed prior to treatment, indicating a fully formed endothelial barrier, and the corresponding results from 1. No treatment added (Black) and 2. Melanoma cell addition (Blue). Created with BioRender.com by Dayna Spurling; Reprinted/adapted with permission from Ref. . 2019, Anchan, A.

Article Snippet: To assess the strength of the cerebral endothelial barrier, hCMVECs were grown on gold-plated 96-well ECIS plates (Applied Biophysics, Troy, NY, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Melanoma Cells Produce Large Vesicular-Bodies That Cause Rapid Disruption of Brain Endothelial Barrier-Integrity and Disassembly of Junctional Proteins

doi: 10.3390/ijms24076082

Figure Lengend Snippet: Human cerebral microvascular endothelial cell CD144 staining after NZM7-S treatment at varied concentrations. hCMVECs were seeded at 20,000 cells per well. NZM7-S were live stained with CytoTrack Red (CTR; red in image) and applied to the apical face of a confluent endothelial monolayer. Cells were co-cultured for 4 h, washed and then fixed. Junctions were stained with anti-CD144 (green). ( A ) An image from the media control well. ( B ) A paired ECIS experiment, where the boxed region shows the point of fixing for the ICC images (4 h). ( C , D,E ) The images of the melanoma-treated wells at different E:T ratios, noted on the left side of the panel. Images were acquired on the Operetta CLS imaging system. Full view images scale bar = 100 µm. Images are representative of 2 independent experiments.

Article Snippet: To assess the strength of the cerebral endothelial barrier, hCMVECs were grown on gold-plated 96-well ECIS plates (Applied Biophysics, Troy, NY, USA).

Techniques: Staining, Cell Culture, Imaging