Fig. 6: AR regulated proliferation is mTOR dependent.

VCaP cells were plated in media supplemented with CFS for 2 days and treated for an additional 48 h with indicated concentrations of R1881 alone or in combination with cycloheximide (CHX). The mRNA expression for PSA (a) and E2F1 (b) were assessed using qRT-PCR. Data are shown as mean ± SD as representative results from three independent experiments, n = 3 technical replicates. c VCaP cells were plated in CFS-supplemented media for 2 days and treated with either vehicle or indicated concentrations of R1881, RTI001 or RU486 for an additional 48 h. Whole cell extracts were probed for RB phosphorylation (pRB), E2F1, FOXM1, PSA, phospho-S6 ribosomal protein (pS6), phospho 4EBP1 (p4EBP1) or β-Actin using western blot. d VCaP cells were treated with indicated concentrations of R1881 alone or in combination with cycloheximide (CHX) for an additional 24 h. Whole cell extracts were probed for RB phosphorylation (pRB), E2F1, FOXM1, PSA, phospho-p70 S6 Kinase (pS6K), phospho-S6 ribosomal protein (pS6), phospho 4EBP1 (p4EBP1) or β-Actin using western blot. e VCaP cells were treated with either vehicle or R1881 (0.06 nM or 10 nM) for indicated time. Whole cell extracts were probed for RB phosphorylation (pRB), E2F1, FOXM1, PSA, phospho-S6 ribosomal protein (pS6), or β-Actin using western blot. f VCaP cells were plated in media supplemented with CFS for 2 days and treated with indicated concentrations of R1881 alone (Veh) or in the presence of 100 nM Torin2 for 48 h. Whole cell extracts were probed for RB phosphorylation (pRB), E2F1, FOXM1, PSA, phospho-p70 S6 Kinase (pS6K), phospho-S6 ribosomal protein (pS6), phospho 4EBP1 (p4EBP1) or β-Actin using western blot. For c–f, representative images are shown from n = 3 independent experiments. Source data are provided as a Source Data file.