Keratinocyte sfm 1

PC3 and LNCaP were purchased from Procell (Wuhan, China), DU145, RWPE-1, and human embryonic kidney 293E (HEK293E) cells from ATCC were gifted by Dr. Qi Li (The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China) and cultured in a humidified environment at 37 °C under 5% CO₂ using their respective media. RWPE-1 cells were maintained in keratinocyte SFM (1×) (Invitrogen, cat. 17,005,042). PC3 and LNCaP cells were cultured in RPMI 1640 supplemented with 15% fetal bovine serum (FBS). DU145 and HEK293E cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 15% FBS. LNCaP and PC3 cells stably overexpressing control and INMT vectors were cultured in RPMI 1640 supplemented with 15% FBS and hygromycin (150 µg/ml) [30 (link), 31 (link)]. LNCaP and PC3 cells stably expressing control shRNA (Sigma-Aldrich, cat. SHC016) and INMT shRNA (Sigma-Aldrich, cat. EHU138831) were cultured in RPMI 1640 supplemented with 15% FBS and puromycin (15 µg/ml) [31 (link)].

The human PCa cell lines 22RV1, LNCaP, DU145, PC-3, VCaP, and normal.
prostate epithelial cells RWPE-1 were purchased from the Shanghai Chinese Academy of Sciences Cell Bank (China). RWPE-1 cells were cultured in Keratinocyte-SFM (1×) (Invitrogen, USA). PC-3, 22Rv1, and LNCaP cells were grown in RPMI-1640 medium (Life Technologies, USA) supplemented with penicillin G (100 U ml^− 1), streptomycin (100 mg ml^− 1) and 10% fetal bovine serum (FBS, Life Technologies, USA). The C4-2B cell line was purchased from the MD Anderson Cancer Center and cultured in T-medium (Invitrogen) supplemented with 10% FBS. DU145 and VCaP cells were grown in Dulbecco’s Modified Eagle’s Medium (Invitrogen, USA) supplemented with 10% FBS [7 (link)].

MEFs and human dermal fibroblasts were cultured in DMEM with 10% FBS, penicillin and streptomycin. Mouse primary keratinocytes were cultured in Keratinocyte-SFM 1× (Invitrogen, 37010–022), 20 μM CaCl₂ and 20 mM HEPES. MCF-10A cells (ATCC CRL 10317) were cultured in DMEM/F12 with 5% horse serum, 20 ng ml⁻¹ EGF, 0.5 μg ml⁻¹ hydrocortisone, 100 ng ml⁻¹ cholera toxin, 10 μg ml⁻¹ insulin, penicillin and streptomycin. MCF-10A cells were used as an epithelial cell model to confirm the findings made with primary cell lines derived from the inducible Pik3ca^H1047R+neo mouse model.