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. Author manuscript; available in PMC: 2018 Apr 9.
Published in final edited form as: Oncogene. 2017 Oct 9;37(5):638–650. doi: 10.1038/onc.2017.371

Figure 5. DHX15, Siah2 and AR form a complex.

Figure 5

A) COS-1 cells were transiently transfected with Flag-vector, Myc-vector, Flag-DHX15, and/or Myc-AR as indicated for 48 h. Cell lysates were immunoprecipitated with antibodies against Flag or Myc. The associated proteins were immunoblotted to detect existence of Flag-DHX15 and Myc-AR in the precipitate. B) LNCaP cells were treated with vehicle or 1nM R1881 for 24 h, Cell lysates were immunoprecipitated with antibodies against AR or DHX15, and the associated proteins were probed with DHX15 and AR antibodies. C) COS-1 cells were transiently co-transfected with Flag-DHX15 and different AR domains as indicated. Cell lysates were immunoprecipitated with antibodies against Flag. The associated proteins were immunoblotted with GFP antibody. D) 293 cells were transfected with Flag-DHX15 and/or HA-Siah2 for 48h. Cell lysates were immunoprecipitated with Flag antibody and subjected to western blotting. E) 293 cells were transfected with Flag-DHX15, GFP-AR and/or HA-Siah2 for 48h, cell lysates were immunoprecipitated with HA antibody and subjected to western blotting and probed with anti-Flag, GFP and HA antibodies. F) 293 cells were transfected with Flag-DHX15-K166A, GFP-AR and/or HA-Siah2 for 48 h, nuclear fractions were extracted and were immunoprecipitated with HA antibody and subjected to western blotting and probed with anti-Flag, GFP and HA antibodies. Data are representative of 3 different experiments.