Bosutinib attenuates phosphorylation of adherens junctions and focal adhesions during Thr-induced endothelial barrier disruption. (A) A schematic overview of the target selectivity of BCR-ABL drugs against 3 kinases that phosphorylate VE-cadherin (Y685) and paxillin (Y118). (B) Thrombin (Thr)-induced temporal phosphorylation kinetics of VE-cadherin (Y685) and paxillin (Y118) were probed using immunoblotting, along with their respective total proteins. GAPDH was used as a loading control (n = 3). (C) Upper panel, Thr-induced fold change of the indicated phosphoprotein in the presence of DMSO vehicle control or bosutinib is plotted above the respective xCELLigence traces. ImageJ quantification of phosphoproteins levels in panel B was adjusted to the total protein amounts, and fold change was calculated by normalizing to the resting state (nontreated, media only). The data are presented as means ± SD (n = 3). Lower panel, quantification of the KI effect on phosphorylation (total protein normalized) in the presence of KIs or DMSO control (100%). The bars represent the median AUC ± SD (n = 3). A 1-way ANOVA, followed by Dunnett multiple comparison test, was used to analyze the data. ∗P < .05. FAK, focal adhesion kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; pVE-Cadh, protein VE-cadherin.