Human umbilical vein endothelial cells huvecs

The structure and characteristics of AXT107 have been previously described.²¹ (link) For the current study, AXT107 and SP2048 were manufactured by solid-state synthesis at Vivitide (Gardner, MA) and amidated on the C-terminal end. AXT107 was labeled on the N-terminus with 5-carboxyfluorescein (FAM-AXT107) and amidated on the C-terminus by GenScript (Piscataway, NJ). Polyclonal rabbit anti-AXT107 antibodies were prepared and purified by Covance Immunology Services (Princeton, NJ). Human umbilical vein endothelial cells (HUVECs) and endothelial growth medium, VascuLife VEGF Endothelial Medium, were purchased from Lifeline Cell Technology (Frederick, MD). Human umbilical vein endothelial cells (HUVECs) stocks were purchased from Lifeline Cell Technology (Frederick, MD) as a ten-donor pool of unspecified sex, to reduce the effects of individual variability on results, and certified to be free of contamination, including mycoplasma. These cells were cultured in VascuLife VEGF Endothelial Medium (Lifeline Cell Technology, Frederick, MD) in a humidified incubator at 37 °C and 5% CO₂.

Chemicals and tissue culture reagents were obtained from Sigma unless otherwise noted. Anhydrous sodium sulfide was purchased from Alfa‐Aesar Inc. Anti‐CD31 antibody was from BD Biosciences (San Jose, CA, USA), and anti-α-SMA antibody was obtained from Sigma-Aldrich. Vectashield plus DAPI was from Vector Laboratories. All secondary fluorophore‐labeled antibodies were obtained from Jackson Immunoresearch Inc (West Grove, PA, USA). Human umbilical vein endothelial cells (HUVECs) were from Lifeline Cell Technology, CA, USA.