Gibco™ DMEM, high glucose

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	supplemented with heat-inactivated 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin (pen/strep)

Protocol tips
supplemented with heat-inactivated 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin (pen/strep)

Publication protocol

The HaCaT cells, a spontaneously transformed nontumorigenic human keratinocyte cell line,16 were routinely cultured in fresh Dulbecco’s Modified Eagle Medium (DMEM; Gibco Laboratories, Grand Island, NY, USA) supplemented with heat-inactivated 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin (pen/strep) at 37 °C in a humidified atmosphere at 5% CO2. Both 3 μg of the circularized HPV-11 DNA and 1 µg of pTK-neo DNA (Invitrogen) were transfected into HaCaT cells on a six-well plate using Lipofectamine™ 2000 reagent (Invitrogen) following the manufacturer’s instructions. After selection with 500 μg/mL G418 (Amersco, Solon, OH, USA) for about one week after transfection and then growing in media containing 250 μg/mL G418, the resulting G418 colonies (HPV11.HaCaT) were pooled as a cell population.

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