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Caspase 3/7 activation assay

NG Nadege Gitego
BA Bogos Agianian
OM Oi Wei Mak
VM Vasantha Kumar MV
EC Emily H. Cheng
EG Evripidis Gavathiotis
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Cells (2.5 × 103 cells/well) were seeded in 384-well white plates overnight and incubated with serial dilutions of Navitoclax (ABT263), A-1331852, ABT199, doxorubicin, BDM19 or vehicle (1% DMSO) using a TECAN D300e Digital Dispenser, in 25 μl of media with 10% FBS. For BDM19, the compound was dispensed in media without FBS and 10% FBS was added after 2 hrs. For the BDM19 and doxorubicin combination, cells were seeded as described above and co-treated with doxorubicin and BDM19 in media without FBS and 10% FBS was added after 2 h. Caspase 3/7 activity was assessed at 2, 6, 8 or 36 hrs as indicated, by the addition of Caspase 3/7 Assay reagents according to the manufacturer’s protocol (Promega), and luminescence measured using a F200 PRO microplate reader (TECAN).

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