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Figure S2. HIP1 R1005E reduces the steady-state nuclear levels of androgen receptor upon androgen stimulation. (A) LNCaP cells transfected with pcDNA3.1 Myc-His6 wild-type HIP1 or HIP1 R1005E were fractionated after Mibolerone or vehicle treatment. Nuclear (N) and cytosolic (C) fractions were resolved by SDS-PAGE (50 µg per lane), transferred to nitrocellulose, and blotted for the AR using a polyclonal antibody (Santa Cruz Biotechnology, Inc.; N20) and a polyclonal antibody against Myc (Cell Signalling) illustrated here with representative blots. (B) The degree of translocation of the AR and HIP1 was quantitated by densitometric analysis of the blots carried out using ImageGauge v4.5 and ImageQuant software. Data shown represents means of five independent experiments +/- SD. (C) Nuclear and cytosolic fractions were also blotted for Lamin B and clathrin as nuclear and cytosolic control proteins.