Figure 1

PRH is hyper-phosphorylated in two prostate cancer cell lines. (a, top) Representative western blot of hypophosphorylated PRH in normal immortalised PNT2-C2 prostate cells and two prostate cancer cell lines; DU145 and PC3. Hypo-PRH was stained with an anti-hypo-PRH mouse polyclonal antibody and Lamin A/C was used as a loading control. (bottom) Densitometric analysis of hypo-PRH levels in three independent experiments. (b, top) Representative western blot of hyper-phosphorylated PRH in the cell lines from a. pPRH was stained with an anti-pPRH rabbit polyclonal antibody and α-Tubulin was used as a loading control. (bottom) Densitometric analysis of pPRH levels in three independent experiments. DU145 *P=0.002, PC3 *P=0.015 (Student’s unpaired t-test). (c) PNT2-C2 cells were treated with either 10% DMSO or 100 μm TBB in DMSO for 72 h. Western blotting was then used to examine the levels of pPRH (as above). Representative of three independent experiments. (d) DU145 cells were treated with (1) 50 μm DMAT, (2) 100 μm DMAT, (3) 50 μm TBB, (4) 100 μm TBB, in DMSO, or (5) 10% DMSO alone for 72 h. Western blotting was then used to examine the levels of pPRH (as above). Representative of three independent experiments.