2.1. HUVEC cell culture and drug treatments

Human umbilical vein endothelial cells (HUVECs) were cultured at 37°C, 5% CO2 in endothelial cell medium (ECM) supplemented with 10% fetal bovine serum, growth supplement, and penicillin/streptomycin all obtained from ScienCell Research Laboratories (Carlsbad, CA). Cells were cultured in 10 cm² plates until they reached ~80% confluence before passaging. Before treatment, cells at 5 to 7 population doublings were plated in the indicated plates for each experiment below at one-half standard density. HUVECs were chronically treated twice over a one-week period with either 0.2% ddH₂O as a vehicle control or 10 μg/mL of the HAART drugs TDF and FTC dissolved in ddH₂O. The vehicle or HAART drugs were added to the ECM before addition to the cells. During the treatment, media was only changed when the second round of treatments occurred which was 3 days after the initial treatment. The control cells were treated with the vehicle in parallel to TDF-FTC treatment. HAART drugs were provided by the NIH AIDS Reagent Program.