Fig. 2.

Androgen-induced CREB activation is AR-dependent. Western blots using phosphorylated (p-CREB) and total (tot-CREB) CREB (43 kDa) antibodies show that exposure to 10 nM DHT did not affect levels of phosphorylated CREB in (A) wild-type (wt) [F (5,11) = 0.02; P = 0.999] and (B) empty vector-transfected (pcDNA-ctl) [F (5,11) = 0.23; P = 0.942] PC12 cells. C, In PC12 cells stably transfected with AR (pcDNA-AR), 10 nM DHT significantly increased CREB phosphorylation [F (5,11) = 6.9; P = 0.004]. D, Hippocampal neuron cultures were pretreated for 2 h with ER antagonist 1 μM ICI 182,780 (ICI), 3β-hydroxysteroid dehydrogenase inhibitor 10 μM trilostane (TRI), or vehicle, followed by exposure to DHT for 2 h. DHT-induced CREB phosphorylation was not blocked by ICI 182,780 or trilostane [F (5,11) = 30.5; P < 0.0001]. Percent phospho-CREB is expressed as a ratio of phosphorylated to total CREB, normalized to the vehicle-treated control condition (bottom panels). Immunoblots are from representative experiments and data show means (± SEM) of the combined experiments (n = 3). *p < 0.05 relative to the vehicle-treated control condition.