Control small interfering si rna

MDA-MB-231 cells were cultured in DMEM (HyClone; Cytiva) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO₂. Subsequently, MDA-MB-231 cells (5×10⁴ cells/well; 24-well plates) were transfected with 100 nM inhibitor control (cat. no. MIH000000; Applied Biological Materials, Inc.), 100 nM miR-1297 inhibitor (cat. no. MIH01244; Applied Biological Materials, Inc.), 1 µg control-plasmid (cat. no. sc-437275; Santa Cruz Biotechnology, Inc.), 1 µg FA2H-plasmid (cat. no. sc-413143-ACT; Santa Cruz Biotechnology, Inc.), 0.2 µM control-small interfering (si)RNA (cat. no. sc-36869; Santa Cruz Biotechnology, Inc.), 0.2 µM FA2H-siRNA (cat. no. sc-93418; Santa Cruz Biotechnology, Inc.), miR-1297 inhibitor + control-siRNA or miR-1297 inhibitor + FA2H-siRNA using Polyplus transfection reagent (Polyplus-transfection SA) according to the manufacturer's protocol. At 48 h post-transfection, cells were used for subsequent experiments.

Control small interfering (si)RNA and two siRNA against DNMT3A were purchased from Santa Cruz Biotechnology and Sigma (Deisenhofen, Germany), respectively. K562DR cells were transiently transfected with either control or DNMT3A siRNA (300 nM) by Amaxa electroporator Nucleofector II (Wako Pure Chemical Industries, Osaka, Japan), using the Nucleofector Kit V (program T-016) as previously described.7 (link)