Sh sy5y neuroblastoma

Human epithelial adenocarcinoma Caco-2 cells (a gift from Dr. Jerrold Turner, University of Chicago) were cultured at 37 °C in Dulbecco’s Modified Eagle Medium (GibcoBRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum and antibiotics. Other cells used were head and neck cancer A431 cells (ATCC, Manassas, VA), skin fibroblasts (a gift from Dr. Edward Schuchman, Mount Sinai School of Medicine), SH-SY5Y neuroblastoma (ATCC, Manassas, VA), and human umbilical vein endothelial cells (HUVECs), whose culture conditions have been reported elsewhere [15 , 26 (link)]. Cells were seeded on gelatin-coated glass coverslips or Transwell® inserts (polyethylene terephthalate, 0.4-μm-pore size; BD Falcon, Franklin Lakes, NJ). The permeability of Caco-2 monolayers was assessed by measuring the transepithelial electrical resistance (TEER), using an EVOM™ volt-ohm meter and STX100 electrodes (World Precision Instruments, Sarasota, FL). Where specified, cells were pre-treated with 10 ng/ml tumor necrosis factor–alpha (TNFα; BD Biosciences, Franklin Lakes, NJ, USA) to mimic an inflammation status [11 ].