Culture and Induction of Reporter Cell Lines

Reporter cell lines that respond to the expression and transcriptional activity of c-Myc (E-H1) or HNF1B (D-D1) derived from NMuMG cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Grand Island, NY, USA) with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA), supplemented with 1% insulin (FUJI FILM Wako Pure Chemical, Osaka, Japan) and 0.5% penicillin-streptomycin (P/S; Invitrogen, Carlsbad, CA, USA) [29 (link)]. The E-H1 and D-D1 cells express c-Myc and HNF1B under the Tet-ON system and the fluorescent protein monomeric Keima (mKeima). Since the ORF of mKeima was inserted after the c-Myc ORF or HNF1B following the internal ribosomal entry site, c-Myc or HNF1B expression could be monitored through the mKeima expression. These proteins were induced by the addition of Doxycycline (DOX, 100 ng/mL) [29 (link)]. The cancer cell lines HeLa, PANC-1, MIA PaCa-2, DU145, and A549 were cultured in DMEM supplemented with 10% FBS and 0.5% P/S, and HCT116 and HL-60 cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco) supplemented with 10% FBS and 0.5% P/S. Cells were incubated at 37°C in a humidified atmosphere containing 5% CO₂.