Figure 2.
Estrogenicity of 8-PN enantiomers compared with estradiol and the standardized spent hop extract. ERα endometrial carcinoma cells (Ishikawa) were grown in 96 well plate (5 × 104 cells/well) in estrogen-free media for 24 h. Subsequently, they were treated with the spent hop extract and compounds at varied concentrations for 96 h at 37 °C, then washed and lysed. Induced estrogen-dependent alkaline phosphatase activity was evaluated spectrophotometrically at 405 nm by the absorbance of the formed p-nitrophenol from the p-nitrophenol phosphate substrate (1 mg/mL). Estrogenic activity of respective treatments was assessed by comparison to the estradiol control (1 nM). The results represent the mean ± SD of at least three independent experiments in triplicate for each sample. The dose response curves were generated using GraphPad Prism.