Transfection

miR-142-5p mimics, miR-142-5p mimics negative controls (NC), miR-142-5p inhibitors, miR-142-2-5p inhibitors negative control (NC), pcDNA3.1 and pcDNA3.1-Bcl-2 associated athanogene 5 (BAG5) were purchased from Shanghai GenePharma Co., Ltd. Cells were seeded in a 6-well plate with a density of 10⁵ cells/ml and pre-incubated for 12 h at 37˚C, before being transfected with 50 nM miR-142-5p mimics or miR-142-5p mimics NC, 2 µg pc DNA3.1 or pcDNA3.1-BAG5, 50 nM miR-142-5p inhibitors or miR-142-5p inhibitors NC with 20 nmol/l Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) then incubated at 37˚C with 5% CO₂ for 48 h. Cells were harvested 48 h after transfection for subsequent experiments. The sequences of the miRNA constructs were as follows: miR-142-5p mimics NC, 5'-GUGUAACACGUCUAUACGCCCA-3'; miR-142-5p mimic, 5'-CAUAAAGUAGAAAGCACUACU-3'; miR-142-5p inhibitors NC, 5'-UCACAACCUCCUAGAAAGAGU-3'; miR-142-5p inhibitors, 5'-AGUAGUGCUUUCUACUUUAUG-3'.