ROCK inhibition induces loss of discontinuous adherens junction. Human umbilical vein endothelial cells were plated at confluence for 48 h and stimulated, as indicated, with 10 ng/ml tumour necrosis factor (TNF)-α in growth medium or TNF-α with 5 μM Y-27632. Cells were then fixed and stained for vascular endothelial (VE)-cadherin, α-catenin and actin filament. Bottom panels show the boxed areas indicated in the merge images at higher magnification: (1) TNF-α, (2) TNF-α + Y-27632. (B) Quantitation of discontinuous junctions in TNF-α-stimulated cells with or without 5 μM Y-27632, +/- standard error of mean of three different experiments. *, P < 0.016.