After transfection for 48 h, A549 and H446 cells were plated into 6-well plates at a density of 0.5–1×103 cells/well; three wells were plated for each experimental condition. The cells were cultured in complete medium supplemented with 30% FBS in an incubator with 37°C for 14 days; the medium was changed every 3 days. During the incubation, the cell state and colony size were observed under a light microscope (magnification, ×100). After culture, cells were washed with PBS, fixed with 4% paraformaldehyde for 20 min at room temperature and stained with 0.2% crystal violet for 5 min at room temperature. The cells were then washed with water, dried, photographed and counted using ImageJ software; >50 cells counted as a colony.

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