Cellular Signaling in Endothelial Cell Regulation

Immortalized human endothelial cells (EAhy926, ATCC) and HEK293 cells were maintained in DMEM supplemented with 10% fetal bovine serum (FBS, Hyclone). Human primary microvascular endothelial cells (HMVEC) were purchased from Lonza and maintained in EGM-2 media with supplements supplied by the vendor; three different batches of primary cells were used in this study as previously described¹⁸ . Primary murine cardiac microvascular endothelial cells were isolated as previously described¹⁹ . Angiotensin II was purchased from Sigma. SNAI2/SLUG promoter-luciferase constructs^{20 (link)}, COL1A2 promoter-luciferase constructs^{21 (link)}, BRG1 expression constructs²² , SLUG expression constructs^{23 (link)}, Sp1 expression constructs^{24 (link)}, and SRF expression constructs^{25 (link)} have been previously described. PFI-3 was purchased from Selleck. Transient transfections were performed with Lipofectamine 2000. Luciferase activities were assayed 24–48 h after transfection using a luciferase reporter assay system (Promega) as previously described^{26 (link)}.