3096 luminometer

For caspase activation assays, cells were plated at a density of 10⁴ cells/well in opaque 96-well plates 24 hours prior to treatment. Twenty-four hours after treatment, activation of effector caspases 3 and 7 was evaluated with Promega’s CaspaseGlo kit (G8090) following manufacturer’s instructions. Resulting mixtures were quantified after 30 minutes of incubation at room temperature in a Beckton Dickinson BD 3096 luminometer.

For caspase 3/7 assays, cells were plated at a density of 10⁴ cells/well in opaque 96-well plates 24 hours prior to treatment. 24 hours after treatment, activation of effector caspases 3 and 7 was evaluated with Promega's CaspaseGlo kit (G8090) as previously described [62 (link)]. Resulting mixtures were quantified after 30 minutes of incubation at room temperature in a Beckton Dickinson BD 3096 luminometer. Data are the average of at least 3 independent experiments performed in triplicates cultures.

For caspase activation assays, cells were plated at a density of 10⁴ cells/well in opaque 96-well plates 24 hours prior to treatment. 24 hours after treatment, activation of effector caspases 3 and 7 was evaluated with Promega's CaspaseGlo kit (G8090) following manufacturer's instructions. Resulting mixtures were quantified after 30 minutes of incubation at room temperature in a Beckton Dickinson BD 3096 luminometer.
For Annexin V/Propidium Iodide citometry assays, cells were seeded in 60-mm culture plates for a final population of 70% confluence. After treatments, cells were collected by trypsinization at the indicated times, pelleted and washed twice with ice-cold PBS. A kit form Immunostep (ANXVF-200T, BB10X-50ML and “PI”) was used for FITC-AnnexinV and Propidium Iodide staining following manufacturer's instructions. Experiments were checked for fluorescence in both green and red channels in a MACSQuant Analyzer 10 cytometer from MACS/Miltenyi Biotec. Dot plot corresponds to data obtained from a representative experiment out of three. Grouped column charts are the average of, at least, 3 independent experiments.