The NCI-H226/SK-MES-1 cells at 1 × 105 cells/well were seeded into 6-well plates in triplicate and transfected with miR-665 mimics, control, miR-665 inhibitor, negative control (NC), TRIM8 siRNA, NC-siRNA, TRIM8 expression plasmid or control vector for 24 h, respectively. These transfected cells were collected by trypsinization and fixed in 70% ice-cold ethanol at 4 °C. Next, 50 μg/mL propidium iodide (PI) and 10 U/mL RNaseA were added in the fixed cells and incubated 20 min for dyeing. The percentages of cell cycle (G1/G0, S and G2/M phases) were detected by using FACSAria flow cytometer (BD Biosciences, USA). Cell cycle was analyzed by using FACSort Cellquect software (BD Biosciences, USA).

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