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Flow cytometry analysis

HX Hongxu Xian
QY Qiaoyun Yang
LX Lin Xiao
HS Han-Ming Shen
YL Yih-Cherng Liou
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Mitochondrial membrane potential was measured by incubating live cells in DMEM supplemented with 10 nM TMRM for 15 min, after which cells were trypsinized and resuspended in PBS. The labeled samples were analyzed using flow cytometry (BD LSR Fortessa or Cytoflex LX) and results were analyzed using SummitTM 4.3 or CyExpert software. Ten thousand events were recorded for each experiment.

For the mt-Keima-based mitophagy assay, WT or Fis1 KO HeLa cells stably expressing mt-Keima were transiently transfected with the plasmid as indicated. Cells were harvested and analyzed for flow cytometry as indicated19.

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