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. 2014 Jun 10;10(6):627–642. doi: 10.7150/ijbs.8756

Figure 6.

Figure 6

AR-Mediated Growth Inhibition does not Require RB, p21 or p27 but does Require DNA Binding. (A) 957E/hTERT-AR cells expressed stable shRNAs to RB. Western blotting documented knockdown of RB protein in 957E/hTERT-Control cells (Control) and 957E/hTERT-AR cells (AR). Actin was used as a loading control. (B) Knockdown of RB failed to override AR-mediate growth inhibition of 957E/hTERT-AR cells (* indicates a p-value < 0.05). (C) Western blotting documented single or double knockdown of p21 and p27 in the same 957E/hTERT-AR cell population; treatment with R1881 failed to produce an elevation of p21 and p27 expression. (D) Clonogenic growth of 957E/hTERT-AR cells in the presence of R1881 documented that individual and double p21 and p27 knockdown failed to prevent growth inhibition by AR (* indicates p-value < 0.05). (E) Western blotting of 957E/hTERT-AR(A573D) cells after treatment with 1nM R1881 for 48 hours. Untreated cells were used as a control (UT). (F) Stable expression of a DNA-binding mutant of AR (A573D) failed to inhibit the clonogenic growth of 957E/hTERT cells in the presence of ligand (+R1881) compared to untreated controls (Untreated).