A549 and SPCA1 were seeded in 12-well plates at a proper concentration and cultured under Nor or CIH condition for 48 h. At the end of CIH cycles, cells were harvested, filtration and centrifugation, and FITC-labeled anti-CD44 (555478) and APC-labeled anti-CD133 (53276) (Cell Signaling Technologies, USA) were used for surface staining. The adherent cells were treated with mitoSOX™ red mitochondrial superoxide indicator (Invitrogen™) at a final concentration of 5 μM for 10 min at 37 °C and washed with PBS three times. Then the cells were harvested, filtration and centrifugation. Cells were detected by the quantitation of fluorescence intensity by flow cytometry. All data were analyzed with FlowJo software (Tree Star Inc., San Carlos, CA).

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