Identification of neuron selective androgen receptor inhibitors

Figure 3 FOXM1 expression correlates with androgen receptor activity. A: Expression of FOXM1 and AR was assessed by Western blot in indicated cell lines; B: GT1-7 cells were transfected with a lentiviral plasmid shRNA targeting FOXM1 as well as luciferase plasmids, treated with drugs listed above, then assessed by luciferase assay. Decreased expression of FOXM1 (RT-qPCR in inset) reduced AR activity and diminished sensitivity to thiostrepton. Fold changes and P values are shown; C: Western blot analysis demonstrates that thiostrepton reduces FOXM1 levels in GT1-7 cells while DHT increases FOXM1 levels; D: GT1-7 cells were treated with the indicated drugs overnight, and nuclear and cytoplasmic fractions were probed by Western blot for the indicated proteins. β-catenin levels in nuclear fraction of thiostrepton treated cells are lower compared to untreated ones. Quantification of β-catenin levels relative to cytoplasmic actin expression is shown; E: β-catenin was immunoprecipitated from GT1-7 cell lysates and Western blot was performed to detect AR and FOXM1. Quantification of AR and FOXM1 levels relative to β-catenin levels in IP is shown. AR: Androgen receptor.