Fig 4. Reengineering of F3 with pure compounds (F3R) does not replicate ERβ agonist activity.

To replicate the desirable estrogenic attribute displayed by F3, ERβ agonism, promoter reporter studies were conducted in HEK293 cells transfected with ERE.vit2.luc and pSG5-hERβ. Cells were induced with 10⁻⁹ M E₂, 9.8 μg/mL F3, 9.8μg/mL F3R (consisting of 1.13 μg mangiferin, 0.39 μg isomangiferin, 1.67 μg scolymoside, 0.02 μg luteolin, 0.01 μg iriflophenone-3-C-glucoside, 0.06 μg p-coumaric acid, 0.06 μg protocatechuic acid as in 9.8 μg F3 (Table 2). Statistical analysis was done using One-way ANOVA with Dunnett’s post-test comparing all columns to either solvent control (black bar) (*, P<0.05; **, P<0.01;***, P<0.001) or F3 (grey bar) (#, P<0.05; ##, P<0.01; ###, P<0.001). The dotted lines through the bars represent the values for solvent control or F3.