Fig. 5.

GT, ABT263, and MK2206 differentially regulate Akt-signaling and AR protein levels. LNCaP cells were treated for 72 h with 1 nM R1881, 10 μM ENZ, or 0.1% DMSO as solvent control. After that, the AR ligands were removed, replaced by fresh medium with 1 μM MK2206 (left panel), 25 nM GT (middle panel), 1 μM ABT263 (right panel), or 0.1% DMSO, and further incubated for additional 24 h (MK2206 and ABT263) or 48 h (GT). Protein extraction was conducted. Detection of AR, Akt, phospho-Akt (p-Akt), S6, and phospho-S6 (p-S6) was performed by Western blotting and normalized to β-Actin levels. For AR, pan-Akt and pan-S6, numbers indicate normalized band intensities relative to DMSO control. For p-Akt and p-S6, upper numbers indicate normalized band intensities relative to DMSO control, while lower numbers indicate ratios of phosphorylated versus total protein levels