Mccoy s 5a medium

Human epithelial ovarian cell lines SK-OV-3, TOV21G, OV90, and CaOV3 were purchased from the American Type Culture Collection (Manassas, VA, USA). SK-OV-3 and CaOV3 were cultured in McCoy’s 5A Medium (Boster, Wuhan, China), while TOV21G and OV90 in mixed MCDB105 Medium (Sigma-Aldrich Co., St Louis, MO, USA) and M199 Medium (Gibco, Invitrogen, Carlsbad, CA, USA) (1:1) supplemented with 10% fetal bovine serum (Gibco, Invitrogen, Carlsbad, CA, USA) and 1% penicillin/streptomycin at 37 °C in a humidified atmosphere of 5% CO₂ and 95% air. The cells were transduced with CMV-Fluc-IRES-RFP lentiviral particles (GeneChem, Shanghai, China). RFP/luciferase-expressing cells were isolated by FACS and used in living imaging. ShDOT1L-1 and shDOT1L-2 shRNA lentiviral particles (GeneChem) were used to knock-down DOT1L expression, targeting 5′-CGGATCTCAAGCTCGCTAT-3′ and 5′-AGTGCTCGAATTGAGAGAA-3′, respectively. ShCON, the short hairpin RNA (shRNA) not targeting any known gene, was used as control. SK-OV-3 and TOV21G cells were transduced with DOT1L shRNA lentiviral particles, or control shRNA lentiviral particles. The cells with stable transfection were selected with puromycin.

Human osteosarcoma cells, U-2 OS, MG-63, and Saos2, as well as human osteoblast hFOB1.19 cells were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human osteosarcoma HOS cells were acquired from the Cell Bank of Wuhan University (Wuhan, China). MG-63 cells were cultured in Minimum Essential Medium (MEM, Gibco, USA), U-2 OS in McCoy’s 5A medium (Boster, Wuhan, China). Saos2 and HOS cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco, USA) containing 1 % penicillin and streptomycin (Gibco, USA), 10 % fetal bovine serum (FBS, Gibco, USA) at 37℃. in a 5 % CO2 humidified incubator HFOB1.19 cells were maintained in MEM containing 10 % FBS, 1 % penicillin/streptomycin, and 0.3 mg/mL G418 (Gibco, USA) at 33.5 °C.