Characterization of Transporter-Expressing HEK293 Cells

Human embryonic kidney (HEK)293 cells (American Type Culture Collection, Manassas, VA, USA) with high expression levels of OATP1B1 (SLC21A6, NM_006446), OATP1B3 (SLC21A8, NM_019844), OATP2B1 (NM_007256), OAT2 (SLC22A7, NM_006672), and OCTN2 (NM_003060) were used. Flp293 cells derived from HEK293 cells stably express the human α1A-adrenoreceptor, with high expression of OAT1 (SLC22A6, NM_004790) and OAT3 (SLC22A8, NM_004254), were also used. HEK293/OATP1B1, HEK293/OATP1B3, HEK293/OATP2B1, HEK293/OAT2, HEK293/OCTN2, Flp293/OAT1, Flp293/OAT3, and mock control cell lines were established by transfecting HEK293 and Flp293 cells with plasmids encoding each transporter. All cells were cultivated in Dulbecco’s Modified Eagle’s medium (FUJIFILM Wako Chemicals, Osaka, Japan) mixed with 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), 1% sodium pyruvate, and 1% penicillin at 37 °C and 5% CO₂.