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. 2015 Jul 24;10(7):e0134015. doi: 10.1371/journal.pone.0134015

Fig 2. FKBP52 Directly Interacts with β-Catenin to Promote Interaction with AR.

Fig 2

(A) In vitro GST-pull down assays were performed with purified, recombinant FKBP52 alone, GST-Tagged β-catenin alone, and both recombinant proteins together. Proteins were visualized on Western Blots with primary antibodies specific to human FKBP52 and β-catenin. (B) A mammalian two-hybrid assay assessing the DHT-dependent activity of a Gal4-mediated luciferase reporter in the presence or absence of a Gal4-AR LBD fusion, Vp16-β-catenin and/or FKBP52 demonstrating that FKBP52 potentiates VP-16-β-catenin/AR interaction in 293 cells. Asterisks (***) denote that hormone-dependent reporter expression in the presence of FKBP52, Vp16-β-catenin, and Gal4-AR LBD was significantly enhanced (p values ranging from < 0.01 to < 0.001) as compared to all other conditions. Hormone-dependent reporter expression in the presence of Gal4-AR LBD with FKBP52 or β-catenin alone did not significantly differ (p > 0.05) from the control with Gal4-AR LBD alone. C. A co-immunoprecipitation to detect β-catenin interaction with FKBP52 and AR with and without DHT and FKBP52 siRNA in LNCaP cell lysates. β-catenin was immunoprecipitated and blots probed for AR, FKBP52 or β-catenin. Inputs are shown at bottom. Note that FKBP52 knockdown results in reduced AR/β-catenin interaction despite similar levels of input.