STAT3 Silencing and Lipid Modulators in Hepatocytes

HepG2 and Huh7 cells (ATCC, Manassas, VA) in culture were treated with 100 nM siRNA-STAT3 (cat: L-003544-00-0005, Dharmacon, Lafayette, CO) or a nontargeting siRNA (siRNA-NTC, cat: D-001810-10-05, Dharmacon) in the presence of Dharmafect transfection reagent for 24 hour. The next day, cells were treated with either combinations of 10 ng/mL IL-1β, 5 μM GW4064 or 10 μM stigmasterol acetate (Stig-Ac) and 10 μM sitosterol acetate (Sit-Ac) (Steraloids, Newport, RI) for 20 hours in DMEM-serum media with transfection mixture, as described.6 (link) In a separate set of experiment, in vitro IL-1β incubation was carried out in HepG2 and in Huh7 cells in culture by treating cells with 10 ng IL-1β (BD Biosciences, San Jose, CA) for 4 hours followed ±5 μM GW4064 overnight in DMEM media.