Rpwe 1

Cell lines LNCaP, PC3, 22Rv1, RPWE-1 and HEK-293T were purchased from ATCC (ATCC, USA). LNCaP and 22Rv1 were maintained in RPMI media (Sigma-Aldrich, USA) supplemented with 10% FBS (Gibco, USA). PC3 was maintained in Ham’s F-12 media (Sigma-Aldrich, USA) supplemented with 10% FBS. RPWE-1 was grown in Keratinocyte Serum Free Medium (ATCC, USA) supplemented with bovine pituitary extract and human recombinant epidermal growth factor. HEK-293T was maintained in DMEM (Sigma-Aldrich, USA) supplemented with 10% FBS. All the cell lines were cultured in a humidified tissue culture chamber with 5% CO₂ at 37°C.

The human prostate cancer cell line PC3 and human lung cancer cell line A549 were purchased from ECACC (Salisbury, UK). The human prostate cancer cell line LNCaP were purchased from ATCC (Manassas, USA). PC3U cells were originally derived from PC3 cells and express more TGFβ receptors than PC3 [45 (link)]. The cancer cell lines were grown in RPMI-1640 with 10% fetal bovine serum (FBS) and L-glutamine (Sigma, St. Louis, MO, USA). The human immortalized normal prostate cell line RPWE1 were from ATCC (Manassas, USA) and cultured in the Keratinocyte Serum-Free Medium with bovine pituitary and human recombinant epidermal growth factor (Gibco-Thermo Scientific, Waltham, MA, USA). The cells were starved 12–18 h in medium supplemented with 1% FBS before EGF, TGFβ (Prospec, Ness-Ziona, Israel), and TNFα (Gibco-Thermo Scientific) stimulation at a concentration of 10 ng/ml. The cell lines were routinely tested for mycoplasma contamination and a panel of pathogens, following the guidelines of Umeå University Animal Care Committee.