Figure 2
From: Validation of a Miniaturized Permeability Assay Compatible with CRISPR-Mediated Genome-Wide Screen
Thrombin increases the paracellular permeability of TIME cell monolayers. (a) The permeability of TIME cell confluent monolayers grown on filter inserts to 2000 kDa FITC-dextran increased in response to treatment by 2.0 U thrombin applied at t = 0 min for the duration of the experiment, as indicated by the augmentation of the fluorescence signal from the bottom wells (mean ± SD, n = 9, **P < 0.01, ***P < 0.001). (b) The impedance of TIME cell monolayers grown on electronic 16-well plates fell sharply after treatment by the indicated dosages of thrombin (applied as above), indicating that the integrity of the monolayer deteriorated. (c) Epifluorescence images of confluent TIME cells in monolayers grown on gelatin-coated coverslips. Intercellular junctions disassembled upon treatment by 2 U/mL thrombin (Thr), as indicated by the change in the VEcad pattern. The cells formed thicker stress fibers and detached from each other. Magnified images of regions surrounded by white frames are shown on the left or right side of each image. The double-pointed white arrow marks a gap between thrombin-treated cells. Scale bars, 50 μm.
