The AR Y267F mutant is defective in androgen- and Ack1-induced nuclear translocation.

<p>(A) COS-7 cells were transfected with the AR expression vector and the constitutively active Ack1 L487F expression vector [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0126270#pone.0126270.ref022" target="_blank">22</a>] and incubated for 48 hrs. The cells were serum-starved overnight and treated with or without DHT (10 nM) for 2 hrs. Subcellular fractionation was performed. Fifteen μgrams of protein from cytoplasmic (C) and nuclear (N) fractions were immunoblotted with the AR antibody. Laminin A/C and 14-3-3 β were used as markers of nuclear and cytoplasmic fractions, respectively. (B) COS-7 cells were transfected with the AR expression vector (full length or truncated) and incubated for 48 hrs. Subcellular fractionation and immunoblotting were performed as described above.