Caspase glo 3 7 activity kit

After incubation with TSA, the caspase 3/7 activity was determined by using a CaspaseGlo 3/7 activity kit (Promega, Madison, WI, USA) according to the manufacturer’s protocol, where 100 µL of the CaspaseGlo 3/7 reagent was added directly to each well (96-well plate). After 60 min of incubation at room temperature, the luminescence was measured with a luminometer (BioTek, Synergy HT, Germany). Each experiment was conducted six times (n = 6).

Caspase 3/7 activity was directly measured at 24, 48 and 72 hours after treatment using Caspase-Glo 3/7 Activity kit (Promega) as per the manufacturer’s protocol. Briefly, the cells (5 × 10⁵/ml) were seeded into 24-well plated and treated with vehicle (DMSO) alone or ruxolitinib at various concentrations for 48 and 72 hours, Caspase-Glo reagent was added and cells were incubated for 1 hour at room temperature in the dark. Relative light intensity was measured in each well using Clarity Luminescence microplate reader (BioTek, Vermont, USA).

Caspase 3/7 activity was assessed using a Caspase-Glo^® 3/7 activity kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions. Briefly, following the pre-determined experimental conditions, H9c2 cells were lysed in 350 µL ice cold lysis buffer. Twenty microliters of the cell lysates was then transferred to a 96-well plate. Thereafter, an equal amount of Caspase-Glo^® reagent (~20 µL) (Promega, Madison, WI, USA) was added to each well and the plate was incubated in the dark. After 30 min, luminescence was measured in a BioTek^® FLX 800 plate reader using Gen 5^® software. Results were normalized to pre-determined protein content using RC DC™ (Promega, Madison, WI, USA) protein assay.