Abstract
Plasma membrane calcium ATPase 1 regulates human umbilical vein endothelial cell angiogenesis and viability
Highlights
Angiogenesis encompasses a series of regulated processes enabling endothelial tip cell migration, stalk cell proliferation and eventual vessel anastomosis and maturation
To determine tubule formation, human umbilical vein endothelial cells (HUVECs) were plated onto GeltrexTM Low Growth Factor Matrix (Invitrogen, 3 × 104 cells per well) in Medium 200 (0.2% FBS) containing either PBS or 25 ng/ml VEGFa-165 (PeproTech)
PMCA1 knockdown leads to impaired HUVEC migration; quantification of wound density and wound width revealed both are significantly reduced in si-PMCA1 HUVECs (Fig. 1B)
Summary
Angiogenesis encompasses a series of regulated processes enabling endothelial tip cell migration, stalk cell proliferation and eventual vessel anastomosis and maturation. Knockdown of ATP2B1 (si-PMCA1), ATP2B4 (si-PMCA4), or nontargeting (si-NT) scrambled control (Dharmacon) in HUVECs (TCS Cellworks) was achieved using published methods [1,2]. HUVECs were plated onto ImageLock 96-well plates (3 × 104 cells per well) and the scratch wound generated using the Incucyte ‘woundmaker’ (Sartorius).
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