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After supernatant treatment for 48h, A549 cells were cultured with DMEM medium in 6-well plates (2×105 cells/mL) for 24 hours. A549 cells were cultured with the prepared conditioned medium for 36 hours. Afterwards, staining buffer (200 μL) was added into A549 cells (1×106 cells/mL). Annexin V (2 μL) was incubated with the cells at 2~8°C in darkness for 15 minutes. Afterwards, PI solution (4 μL) was added for incubation at 2~8°C in darkness for 5 minutes. A549 cell apoptosis was detected by a flow cytometer (BD, Franklin Lake, New Jersey, USA).
Copyright and License information: ©2020 Yu et al.
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