Hek293t cell line

The human melanoma cell lines IRNE and mel1359 used were kindly provided by G. Pawelec (University of Tuebingen, Germany) and have recently been published.¹⁸ These cell lines were maintained in Roswell Park Memorial Institute 1640 Medium (RPMI, Invitrogen) supplemented with 10% (v/v) foetal bovine serum (FCS) (PAA, Pashing, Austria), .1 mM non‐essential amino acids (Gibco, Dublin, Ireland), 2 mM L‐glutamine (Lonza, Basel, Switzerland), and 1% penicillin/streptomycin (v/v; PAA) at 37°C, 5% CO₂. The HEK293T cell line was acquired from DSMZ and was cultured in DMEM medium (Sigma‐Aldrich, Munich, Germany) supplemented with 10% (v/v) FCS (PAA), .1 mM non‐essential amino acids (Gibco), 2 mM L‐glutamine (Lonza) and 1% penicillin/streptomycin (v/v; PAA) at 37°C, 5% CO₂.

The HEK293T cell line was purchased from Sigma (Cat. No.: 12022001-1VL). Cultivation medium DMEM, FBS, and supplements were purchased from Sigma Aldrich. Radiolabeled 14C uric acid MC-1394 was purchased from Hartmann analytic GmbH, Braunschweig, Germany. Plasmids with cloned SLC22A6 WT (wild type) and SLC22A8 WT pLenti-C-mGFP-P2A-Puro and mGFP primary antibody OTI2FG were purchased from Origene, Rockville, MD, USA. Mutagenesis was accomplished using Geneart Site-directed mutagenesis kits from Qiagen (Cat. No.: A13282), Hilden, Germany. Beta-actinin primary antibody was purchased from Cell Signaling (clone 8H10D10), Danvers, MA, USA. Rabbit anti-mouse HRP conjugated secondary antibody was provided by Invitrogen, cat. No.: A90-117P. Kits for plasmid DNA isolation were purchased from Qiagen, Hilden, Germany. Bradford assay kits were purchased from Biorad, Hercules, CA, USA. Cultivation plastic and were provided by VWR, Radnor, PA, USA. PVDF blotting membranes were purchased from Merck Immobilon (Cat. No. IPVH 07850). Other common chemicals came from Penta chemicals, Praha, Czech Republic or Sigma Aldrich, St. Louis, MO, USA.