Six well tissue culture plate

For fluid shear stress, cells were maintained in DMEM containing 1% serum and 10 µg/mL heparin on six-well tissue culture plates (Costar) for 24 h before initiating shear stress experiments. A cone-plate viscometer was designed and built such that it accepts six-well plates.^{27 (link),28} This allowed the PAEC monolayer to be subjected to a radially constant fluid shear stress at laminar flow rates representing levels of shear stress within physiologic parameters. Typical physiologic shear stress in the major human arteries is in the range of 5–20 dynes/cm².^{33 (link)} Thus, we imparted a shear stress of 20 dynes/cm² for 8 h to mimic the upper limit of the physiologic range. For cyclic stretch, cells were maintained in DMEM containing 1% serum and 10 µg/mL heparin on six-well BioFlex plates coated with collagen type I (FlexCell) for 24 h, then subjected to biaxial cyclic stretch using the FlexCell 3000 Strain Unit. Plates were placed on a loading station and stretched by applying an oscillatory vacuum to the underside of the membranes. Cells were stretched at a frequency of 1 Hz with 20% amplitude for 8 h in accordance with previous studies.^{34 (link)}