Phosphorylation site mutants of AR are defective in stimulating cell proliferation and anchorage-independent soft agar colony growth.

<p>(A, B) LNCaP cells stably overexpressing full length AR (A) or truncated AR (B) were grown in a 96-well plate for 72 hours in media with charcoal-stripped serum and 0, 0.1, 1, or 10 nM of DHT, as indicated. Absorbance at 450 nm was measured after incubation with the colorimetric dye WST-8 for 4 hours. Values were normalized to vector control cells treated with 0 nM DHT. Data shown are the mean <u>+</u> standard deviation of three independent experiments, each performed with triplicate or quadruplicate wells. *, p<0.001 vs AR-WT; **, p<0.05 vs AR-WT by two-group t-test adjusted using the Bonferroni method. (C, D) Cells stably overexpressing full length AR (C) or truncated AR (D) were plated in soft agar and incubated for 3 weeks. Colonies were stained and counted. A representative view of the colonies is shown below. Data shown are the mean <u>+</u> standard deviation of two (C) or three (D) independent experiments with triplicate samples. *, p<0.001 vs AR-WT; **, p<0.05 vs AR-WT by two-group t-test adjusted using the Bonferroni method.