We have previously established stable PC-3 and DU 145 human PCa cell lines that express the wild-type KLF5, the Ac-KLF5-mimicking mutant KLF5K369Q (KQ), the acetylation-deficient mutant KLF5K369R (KR), and the pLHCX vector control [22 (link)]. PC-3 cells expressing different forms of KLF5, including PC-3-KLF5, PC-3-KQ, PC-3-KR, and PC-3-pLHCX, were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS, Biological Industries, HAMEK, Israel) and 1% penicillin/streptomycin (100 U/mL, Biological Industries). DU 145 cells expressing different forms of KLF5 were cultured in the minimum Eagle’s medium (MEM) (Corning, New York, USA) with 10% FBS.
PC-3-KLF5, PC-3-KQ, and PC-3-KR cells were infected with lentiviruses expressing the luciferase (HBLV-LUC-PURO, Hanbio, Shanghai, China). Infected cells were selected for stable cell populations in a medium containing puromycin (2 μg/mL).
RAW264.7 macrophage cell line was obtained from the American Type Cell Culture (ATCC, Manassas, VA, USA) and cultured in Dulbecco's modified eagle medium (DMEM) (Biological Industries) supplemented with 10% FBS. All cells were cultured at 37°C in a humidified atmosphere with 5% CO2.
PC-3-KLF5, PC-3-KQ, and PC-3-KR cells were infected with lentiviruses expressing the luciferase (HBLV-LUC-PURO, Hanbio, Shanghai, China). Infected cells were selected for stable cell populations in a medium containing puromycin (2 μg/mL).
RAW264.7 macrophage cell line was obtained from the American Type Cell Culture (ATCC, Manassas, VA, USA) and cultured in Dulbecco's modified eagle medium (DMEM) (Biological Industries) supplemented with 10% FBS. All cells were cultured at 37°C in a humidified atmosphere with 5% CO2.
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