Following transfection, A549 and H446 cells were plated into 96-well plates at a density of 2×103 cells/well and cultured in DMEM supplemented with 10% FBS. For certain experiments, cells were also treated with shikonin [0, 10, 20 or 50 µM] for 24, 48, 72 or 96 h at 37°C. At each time point, Cell Counting Kit-8 (CCK-8) reagent (Dojindo Molecular Technologies, Inc.) was added to each well and incubated for 2 h at 37°C. The optical density (OD) value was measured at a wavelength of 450 nm using a microplate reader (Thermo Fisher Scientific). The experiment was repeated in triplicate.

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