Cell Line Characterization and Proliferation Assays

Cell lines DU145 (ACC 261), HEK293 (ACC 305), HeLa (ACC 57), IPC298 (ACC 251), IGR39 (ACC 239), IMR32 (ACC 165), and SH-SY5Y (ACC 209) were purchased from DSMZ (Braunschweig, Germany). MDA-MB-435S (HTB129) cells were from ATCC (Manassas, VA), PNT2 cells (ECACC95012613) were obtained from ECACC (Salisbury, UK). GL15 cells were kindly provided by Dr. Fioretti (University of Perugia, Italy). Each cell line was cultured in their respective recommended medium supplemented with 10% FCS (PAA Laboratories) at 37 °C in humidified 5% CO₂ atmosphere. For stablly transfected cell lines (HEK expressing K_V10.1 in the pTracerCMV vector, a cell line routinely used in our laboratory (4 (link), 6 (link)– (link)8 (link)), the selection compound Zeocin (Calya) was added to the culture medium at 3 μg/ml. Transient transfections were performed using FuGENE (Roche Applied Science) or Lipofectamine 2000 (Invitrogen). Proliferation was estimated using Alamar Blue (BIOSOURCE) or WST assays (Roche Applied Science) as described (50 (link)) or by live cell imaging in an IncuCyte Zoom system (Essen Biosciences) to determine the percent confluence as a function of time.