Fig 2: ER and GPER1 are essential for isoflavone-augmented dendrite arborization in Purkinje cells. Cerebellar cells were cultured for 17 days, followed by an immunocytochemistry analysis of Purkinje cells with Calbindin D-28K (red) and DAPI (blue). (A) Representative photomicrographs showing the co-exposure effects of genistein with ICI 182,780 or G15 on the morphology of Purkinje cells. (B) Changes in the dendritic areas in Purkinje cells following S-equol, G15, and/or ICI 182,780 treatment (n = 15). (C) Representative photomicrographs showing the effects of genistein on the morphology of Purkinje cells after ERa, ERß, or GPER1 knockdown. Quantitative analysis of the effects of isoflavones after exposure to ERa siRNA (n = 15) (D), ERß siRNA (n = 15) (E), siRNA against ERa and ERß (n = 15) (F), and GPER1 siRNA (n = 15) (G) on the dendritic areas in Purkinje cells following isoflavone treatment. Bars indicate 50 µm. Data are shown as the mean ± standard error of the mean and represent at least three independent experiments. #### p < 0.0001 indicates statistical significance according to two-way or one-way analysis of variance, followed by the post hoc Tukey test compared with E2 (10 nM). **** p < 0.0001 and *** p < 0.001 indicate statistical significance according to two-way or one-way analysis of variance, followed by the post hoc Tukey test.