AM966, an Antagonist of Lysophosphatidic Acid Receptor 1, Increases Lung Microvascular Endothelial Permeability through Activation of Rho Signaling Pathway and Phosphorylation of VE-Cadherin

Figure 1

AM966 increases permeability in HLMVECs. (a) Confluent HLMVECs were plated on gold electrodes and TEER changes were monitored in real time using ECIS. After baseline resistance was stable, DMSO, AM966 (1.0 μM), LPA (5 μM), or AM966 + LPA was added to each well. The TEER tracing represents pooled data (±SEM) from 3 independent experiments. (b) The electrical resistance during indicated time period (a) was quantified and statistical analysis was performed. (c) Confluent HLMVECs were plated on gold electrodes and TEER changes were monitored in real time using ECIS. After baseline resistance was stable, different doses of AM966 (0.1, 1.0, or 10.0 μM) were added to each well. The TEER tracing represents pooled data (±SEM) from 3 independent experiments. (d) The resistance in response to AM966 treatments during indicated time period (c) was quantified and statistical analysis was performed. (e) HLMVECs (~100% confluence) were grown on a glass bottom coverslip and serum deprived for 3 h, and then the cells were treated with DMSO or AM966 (1 μM) for 30 min. Immunofluorescence staining of VE-cadherin (green), F-actin (red), and nuclei (blue) was examined by a Zeiss LSM 510 confocal microscope. Scale, 15 μm. Paracellular gaps are marked by arrows. Shown are representative images from three independent experiments.

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