Proper functioning of intracellular membrane trafficking in endothelial cells is indispensable for maintaining endothelial barrier, as well as for regulated secretion of a variety of pro- and anticoagulants and inflammatory proteins. While the involvement of several heterotrimeric G proteins in regulation of membrane trafficking has been documented, there is only one work published on the possible role of Gα12 in this respect in PC12 cells, where Gα12 has been reported to inhibit exocytosis. In the present study, we addressed possible roles of Gα12 in membrane trafficking in endothelial cells, using human umbilical vein endothelial cells (HUVEC) as a model. Using confocal microscopy, ECIS measurements, and velocity sedimentation assays, we examined the effects of overexpression of wild-type or constitutively active Gα12, as well as siRNA-mediated depletion of endogenous Gα12, on several markers in resting or thrombin-stimulated HUVEC. Our data indicate that Gα12 has stimulatory effect on exocytosis in HUVEC. Moreover, we were able to detect macromolecular complexes of Gα12 with transported proteins, suggesting its direct involvement in membrane trafficking.
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