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The cell suspension was passed 20 times through a 25 G needle, and nuclei and unbroken cells were removed by centrifugation at 700 g for 10 min at 4 C
For perfusion assays with HeLa cells, NAPstar3b was monitored using the Ex and Em settings as above, HyPer7 was monitored using- Ex: 405 20 nm, Em: 525 50 nm and Ex: 470 40 nm, Em: 525 50 nm
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