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 3

AM966 induces phosphorylation of VE-cadherin. (a) Confluent HLMVECs were treated with AM966 (1.0 μM) or LPA (5.0 μM) for 30 min after 3 h starvation. Cell lysates were immunoblotted with phospho-VE-cadherin (P-VE-cadherin), total VE-cadherin, phospho-Erk1/2 (P-Erk1/2), and total Erk1/2 antibodies. (b) Analysis of P-VE-cadherin and P-Erk1/2 by densitometry of the results in (a) was performed by Image J software (), and statistical analysis was shown. (c) Confluent HLMVECs were treated with AM966 (1.0 μM) for indicated time after 3 h starvation. Cell lysates were immunoblotted with P-VE-cadherin and total VE-cadherin antibodies. (d) Analysis of P-VE-cadherin by densitometry of the results in (c) was performed by Image J software (), and statistical analysis was shown. (e) Confluent HLMVECs were treated with AM966 (0 to 10.0 μM) for 30 min after 3 h starvation. Cell lysates were immunoblotted with P-VE-cadherin and total VE-cadherin antibodies. (f) Analysis of P-VE-cadherin by densitometry of the results in (e) was performed by Image J software (), and statistical analysis was shown. Shown are representative blots from three independent experiments.

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