Figure 5.

The impact of clotrimazole and fenticonazole on ERα transcriptional activation. Western blotting and analysis of the ERα and the ERα phosphorylation status on S residue 118 (pS118) induced by 17β-estradiol (E2 1 nM–30 min) in MCF-7 cells pre-treated with clotrimazole (Clo 10 µM) (a) and fenticonazole (Fenti 10 µM) (b) for 72 h. (c) Densitometric analysis is relative to panel (a,b). The loading control was done by evaluating vinculin expression in the same filter. Panels show representative blots of three independent experiments. Significant differences with respect to-sample are calculated by Student t-test and indicated by **** (p-value < 0.0001). Significant differences with respect to the E2 sample are calculated by Student t-test and indicated by °°°° (p-value < 0.0001). (d) Estrogen response element promoter activity in MCF-7 ERENLuc cells pre-treated with clotrimazole (Clo 10 µM) and fenticonazole (Fenti 10 µM) for 72 h and then treated with 17β-estradiol (E2 1nM) for an additional 24 h. (e) Estrogen response element promoter activity in Y537S ERENLuc cells treated with fulvestrant (ICI-100nM), clotrimazole (Clo), and fenticonazole (Fenti) at the indicated doses for 72 h. The experiments were performed twice in duplicate. Significant differences with respect to untreated (i.e., -,-,- or 0) sample are calculated by Student t-test and indicated by **** (p-value < 0.0001). Significant differences with respect to the E2 sample are calculated by Student t-test and indicated by °°°° (p-value < 0.0001). Significant differences with respect to Clo- or Fenti-alone treated sample are calculated by Student t-test and indicated by ^^^^ (p-value < 0.0001).