Hoechst 33342 and AO/EB staining were used to analyze the morphological characteristics of apoptosis of A549 cells treated with CK and CK-Ngs. The A549 cells were cultured overnight in 2 mL of culture medium at a density of 2 × 105 cells/well in 6-well plates. Then, CK and CK-Ngs were added to the cells for 96 h. Control group treated with culture medium. After the supernatant was removed and cells were washed 3 times with PBS, the cells were stained with 10 μg/mL Hoechst 33342 solution for 15 min in the dark or with AOEB staining solution for 10 min, and still washed three times with PBS. The stained cells were observed with a fluorescence microscope (Nikon, Tokyo, Japan).

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