Palbociclib

LNCaP, HEK293T, VCaP, and PC-3 cells were obtained from the American Type Culture Collection (ATCC). C4-2 cells were purchased from Uro Corporation. LNCaP-RF cells were described previously (14 (link)). HEK293T cells were maintained in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS). VCaP cells were maintained in DMEM supplemented with 13% FBS. C4-2, LNCaP, LNCaP-RF, and PC-3 cells were maintained in RPMI 1640 medium supplemented with 10% FBS. All cell lines were authenticated (karyotyping, mutations in p53 and ERG fusions, and AR, PTEN, p53, and ERG protein expression) and used within 6 months of thawing. No mycoplasma contamination was detected in these cell lines by testing with the Lookout Mycoplasma PCR Detection Kit (Sigma-Aldrich). Charcoal-stripped serum (CSS) was purchased from Thermo Fisher Scientific-Gibco (#12676029). Enzalutamide was kindly provided by Medivation. LNCaP-RF cells were treated with 10 μM of enzalutamide for 72 hours unless otherwise noted. Palbociclib (PD-0330991) was obtained from ApexBio. LNCaP-RF cells were treated with 1 μM of Palbociclib for 72 hours unless otherwise noted. For combination treatment, LNCaP-RF cells were treated with 10 μM enzalutamide and 1 μM Palbociclib for 72 hours.