Tpt chloride

Unless otherwise stated, all chemicals were dissolved in DMSO at 0.05% (v/v). Control groups were exposed to DMSO at 0.05% (v/v). Cell viability was evaluated using an MTT assay as described previously (Pu et al., 2019 (link)). Theca cells were seeded, cultured in growth medium, and exposed to TPT chloride (Cat# 45492, Sigma, St. Louis, MO, USA; concentrations: 0, 0.1, 1, 10, 20, 50, 100 and 1,000 ng/ml) and/or an LXR antagonist (GSK2033; Cat# SML1617, >98%, Sigma, St. Louis, MO, USA; concentrations: 0, 0.1, 0.5, 1, 2 and 5 μM) and RXR antagonist (HX531; Cat# SML2170, >98%, Sigma, St. Louis, MO, USA; concentrations: 0, 0.1, 0.5, 1, 2 and 5 μM) and PPARγ antagonist (T0070907; Cat# T8703, >98%, Sigma, St. Louis, MO, USA; concentrations: 0, 0.1, 0.5, 1, 2.5, 5, and 10 μM) in the growth medium for 72 h. Cells were then incubated with MTT (50 μg/ml) for 4 h in a phenol red free DMEM medium. After replacing with DMSO, cells were vortexed for 10 min, and cell viability measured by absorbance quantification at 570 nm with a microplate reader (SpectraMax M5e, Molecular Devices LLC, Sunnyvale, CA, USA). Chemical exposure was begun right after serum starvation for 48 h on pre-luteinized and 72 h on cells during their luteinization.