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To analyze HARA and HARA-B4 cell death due to a loss of adherence, the two cell types were stimulated and then seeded into low-adhesive plates at a density of 10 × 105 cells per well. After 1, 4, or 7 days, the cells in each group were harvested and centrifuged at 1,000 rpm for 5 min; after which, they were washed with PBS, and then stained with Annexin V (Invitrogen, Carlsbad, CA, USA) solution and propidium iodide (PI) solution according to the instructions of the manufacturer. The percentage of cells with anoikis was analyzed by flow cytometry.
Copyright and License information: ©2021 She, Yang, Li, Xiong and Zhou
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