FIG.5.
AR acetylation mutants convey contact-independent growth. (A and B) MTT assay of DU145 stable cell lines expressing either pcDNA3, wild-type AR, or AR acetylation site mutants. Equal numbers of cells were seeded into 96-well plates, treated with either DHT, TSA, or SAHA for 24 h, and the MTT assay was conducted, measuring absorbance at 570 nm. (C and D) DU145 cells stably expressing wild-type AR or AR acetylation site mutants were seeded in soft agar. Phase contrast image of the colonies from a representative experiment is shown (magnification, ×100). Colony numbers and size (percentage of colonies with >100 cells) are shown at day 14. (E and F) Nude mice were implanted with 106 cells of stable lines expressing either wild-type AR or AR acetylation site mutants. The mean volume of DU145 tumors grown in nude mice were shown at each time point. (G and H) Apoptosis in implanted tumors was assessed by TUNEL staining for wild-type AR, ARK630Q, and ARK630T (n = 4).