Hepg2 cell

Human hepatocellular carcinoma HepG2 cells and mouse neuroblastoma Neuro2a cells were purchased from ATCC (Rockville, MD, USA). Control human fibroblasts were obtained from Coriell Institute for Medical Research (Camden, NJ, USA). HepG2 and human dermal fibroblasts were maintained in DMEM with 10% fetal bovine serum and Neuro2a were maintained in EMEM with 10% fetal bovine serum. To determine the effect of drugs, cells were plated in 96-well plates and incubated at 37 °C in 5% CO₂ for 48 h in presence and absence of different concentrations of ARI, TRZ, and ARI + TRZ. The treatment was performed in cholesterol deficient medium. HepG2 and human fibroblast cultures were grown in DMEM with 10% delipidated fetal bovine serum, and Neuro2a were grown in EMEM plus N2 supplement. At the end point of the incubation, Hoechst dye was added to all wells in the 96-well plate, and the total number of cells was counted using an ImageXpress Pico and cell counting algorithm using CellReporterXpress. After removing the medium, wells were rinsed twice with 1× PBS and then stored at −80 °C for sterol analysis. All samples were analyzed within 2 weeks of freezing.

Human pancreatic cancer cell lines PANC-1, AsPC-1, MIA PaCa-2, BXPC-3, liver cancer HepG2 cells, colon carcinoma LS-180 cells, and cervical carcinoma HeLa cells were purchased from ATCC (Manassas, VA), and human hepatocellular carcinoma Huh-7 cells were bought from Riken Cell Bank (Wako, Saitama, Japan). All cells were maintained in Dulbecco's Modified Eagle Medium (DMEM) or Eagle's Minimum Essential Medium (EMEM) containing 10% FBS, 100 U/ml of penicillin sodium, and 100 μg/ml of streptomycin sulfate at 37°C in a humidified atmosphere of 5% CO₂. The miR-1291-expressing and control PANC-1 cells were established recently in our lab [30 (link)]. AsPC-1 cells stably transfected with miR-1291 expression plasmid and empty control vector were developed in the same manner.

HepG2 cells were purchased from ATCC (Virginia) and cultured in DMEM plus 10% FBS. pGL3-PPARγ2 (−2.5 kb) plasmid DNA and plasmids expressing HNF4α, SHP, or β-galactosidase (β-gal) were transfected into HepG2 cells. After 36 h, luciferase activity was determined and normalized to β-gal activity.

We cultured K562 cells (ATCC) in IMDM (Gibco #12440) supplemented with 10% fetal bovine serum (FBS) and 1% pen/strep and maintained in a humidified chamber at 37 °C with 5% CO₂. We cultured HepG2 cells (ATCC) in EMEM (ATCC #30-2003) supplemented with 10% FBS and 1% pen/strep, maintained in a humidified chamber at 37 °C with 5% CO₂.