Figure 2.
2-ME down-regulates AR, required for G2/M phase arrest. (a,b) Western blot for AR and GAPDH (as loading control) in lysates from (a) LNCaP and (b) C4-2 cells treated for 24 h with the indicated concentrations of 2-ME. (c,d) qRT–PCR of mRNA at indicated concentrations of 2-ME in LNCaP (c) and C4-2 cells (d). AR mRNA levels were plotted after normalizing to that of GAPDH. FACS analysis of (e) LNCaP and (f) C4-2 cells depleted of AR by siRNA, compared with control siRNA treated cells (left half). Results are representative of at least three independent experiments. (g) PC3-M cells stably infected with either empty lentiviral vector or lentivirus expressing AR were treated without or with 2-ME for 24 h. FACS analysis of PC3-M stably expressing empty vector or WT-AR (left) and immunoblot for AR in PC3-M cells expressing AR (5 μg of protein, right), with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) serving as the loading control.