Figure 6.

Physiological outcomes of D in mixture with its metabolites. (a) Analyses of cellular DNA content obtained from PI assay, Western blot (left) and densitometric analyses (right) of NGB levels (b) in MCF-7 cells. (c,d) Western blot (up) and densitometric analyses (bottom) of PARP-1 cleavage in MCF-7 cells. The MCF-7 cells were treated for 24 h with either vehicle (DMSO) or E2 (10 nM) or D, D4S or Eq (1 µM); some samples were treated with all the sulfate metabolites (D4S, D7S and DDS, 1 µM each) or all the gut metabolites (Eq and O-DMA, 1 µM each) in presence or absence of D (1 µM). Data are mean ± SD of three different experiments. (*) p < 0.001 was calculated with Student t test versus vehicle in (a). In b, data are the mean ± SD of five different experiments: p < 0.001 was determined with Student t test with respect to the vehicle (*) or to D-treated samples co-stimulated with metabolites (°). In (c,d), data are the mean ± SD of three different experiments. p < 0.001 was determined with Student t test with respect to the vehicle (*) or Pacl-treated (°) or E2 untreated samples co-stimulated with the metabolites (+). E2: estradiol; NGB: neuroglobin; DMSO: dimethyl sulfoxide; D: daidzein; Eq: equol; O-DMA: o-desmethylangolesin; D7S: daidzein-7-sulfate; D4S: daidzein-4′-sulfate; DDS: daidzein-7,4′-disulfate; Pacl: paclitaxel.