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. 2019 Oct 18;20(20):5178. doi: 10.3390/ijms20205178

Figure 3.

Figure 3

Effects of S-equol on mouse cerebellar astrocyte proliferation. Mouse primary cerebellar astrocytes were cultured for seven days followed by a BrdU incorporation assay, MTS cell proliferation assay, and immunohistochemical analysis with pERK1/2 and DAPI staining. (A) Representative photomicrographs showing the effects of S-equol on the BrdU incorporation assay. The right panel shows the changes in the percentages of BrdU positive cells after exposure. (B) Time-dependent changes in the effect of S-equol, G15, and/or ICI on cellular proliferation. Astrocytes were exposed to S-equol, G15, and/or ICI for 24, 48, and 96 h, respectively. Cell viability was determined with an MTS assay and the number of viable cells was calculated as a percentage of the control viability. (C) Representative photomicrographs showing the immunohistochemistry for pERK1/2 and DAPI staining to examine the effect of S-equol, G15, and/or ICI on pERK1/2 expression. The right panel shows the changes in the percentages of pERK1/2 positive cells after exposure. The bars indicate 50 μm. Data are expressed as the mean ± SEM (n = 50 determinations) and are representative of at least three independent experiments. ****p < 0.0001, indicates statistical significance according to Bonferroni’s test compared with control (-). ####p < 0.0001, ###p < 0.001, and ##p < 0.005 indicate statistical significance according to Bonferroni’s test compared with S-equol.