Gw7647

Day 10 WT or ERRα/γ KO hiPSC-CMs were cultured with the previously reported maturation cocktail^{15 (link)} for 7 days with minor modification as described below. CDM3 media was supplemented with 4 nM 3,3′,5-triiodo-L-thyronine (Sigma-Aldrich, T5516-1MG), 100 ng/mL dexamethasone (Sigma-Aldrich, D4902-25MG), 1 μM GW7647 (Cayman, 10008613), 200 μM palmitate (P9767, Sigma-Aldrich)-conjugated with fatty acid-free bovine serum albumin (Sigma-Aldrich, A6003-100G), and 500 μM carnitine (C0283-5G, Sigma-Aldrich).

Tg(fabp10a:pt-β-catenin) larvae were treated with 0.1% dimethyl sulfoxide (DMSO), 1 µM GW7647 (Cayman Chemical, Ann Arbor, MI), 2 µM K-975 (MedChemExpress, Princeton, NJ), 15 µM Etomoxir (Cayman Chemical, Ann Arbor, MI), 2 µM AG1478 (ApexBio, Houston, TX), or 7 μM CAY10599 (Cayman Chemical, Ann Arbor, MI) for 48 h from 12 or 13 days post-fertilization (dpf). Compounds were refreshed every 24 h. The larvae were fed prior to the drug treatments but fasted during the treatments. They were harvested at 14 or 15 dpf for subsequent analyses.

The human intestinal epithelial cell lines HT–29 was obtained from the American Type Culture Collection (ATCC, Rockville, MD). HT–29 cells were cultured in DMEM supplemented with 10% heat-inactivated fetal calf serum (FCS), 2 mM L-glutamine (Sigma), 1X Non essential Amino acid (Invitrogen), penicillin (50 IU/ml) and streptomycin (50 μg/ml) in an humidified atmosphere containing 10% CO2 at 37°C. After seeding, cells were grown 48h in 6 or 12 wells plate in antibiotic-free medium at 3.25X10⁵ and 6.5X10⁵ respectively. Medium was changed just before the addition of bacterial or reagents for 6h.
Rosiglitazone (used as positive control), GW9662, GW6471 and GW7647 (Cayman chemicals) were dissolved in DMSO following the manufacturer’s instructions and diluted at 100μM in antibiotic-free DMEM. They were used at a final concentration of 10μM except for GW6471 at 1μM. The antagonists (GW9662, GW6471) were added 1h before challenging with rosiglitazone or GW7647 respectively.