MOESM14 of MicroRNA-155 contributes to shear-resistant leukocyte adhesion to human brain endothelium in vitro
mediaposted on 10.10.2019, 08:55 by Camilla Cerutti, Patricia Soblechero-Martin, Dongsheng Wu, Miguel Lopez-Ramirez, Helga Vries, Basil Sharrack, David Male, Ignacio Romero
Additional file 14: Video S13. Shear-resistant firm adhesion of JURKAT cells to CONTROL Scrambled-Anti-miR transfected and stimulated with TNFα and IFNγ hCMECD3 cells. JURKAT cells were pulled through the channel over hCMEC/D3 monolayers under low shear (0.5 dyn/cm2). After 5 min, flow shear stress was increased (1.5 dyn/cm2) to challenge non-firmly adherent JURKAT. The number of arrested JURKAT constantly increased during the accumulation phase and only the one that remained stationary on the endothelial monolayer were manually counted in 5 different FOV along the channel of Ibidi m-Slide VI. FITC channel was used to count fluorescently firmly adhered Jurkat cells. Objective 10x, at 40 images per min (1 phase-contrast and 1 FITC), field of view 640 μm × 480 μm, recording time 5.5 min.