Caspase glo 8 or 9 assay kit

The cells were seeded in 12-well plate (1 × 10⁵ cells/well) and treated with yamogenin at concentrations of 10.0–70.0 µg/mL. The concentration of ethanol added to the cells did not exceed 0.7% (v/v). After 24 h of treatment, caspase-3/7 activation was measured, and an estimation of the apoptotic status of the cells was conducted. The cells were stained with a fluorescent reagent that contained a DNA-binding dye linked to a DEVD peptide substrate. The dye is released from the complex when caspase-3/7 is active. A cell marker (7-AAD) was also used in the assay as a marker of dead cells. The cells were analyzed with flow cytometry (Muse Cell Analyzer). The experiments were performed in three independent repeats.
The caspases-8 and -9 activity level in the cells was determined with Caspase-Glo 8 or 9 Assay Kit (Promega, Madison, WI, USA) and Glomax Multi + Detection System (Promega). The cells were seeded in 96 well plates (1 × 10⁴ cells/well), and after 24 h of incubation, they were treated with yamogenin at concentrations of 10–70 µg/mL for 24 h. The experiments were performed in three independent repeats.

The caspase-8/9 activity level in the cells was determined with Caspase-Glo 8 or 9 Assay Kit (Promega, Madison, WI, USA) and Glomax Multi+ Detection System (Promega, Madison, WI, USA) according to the manufacturer’s instruction. The cells were seeded in 96-well plates (1 × 10⁴ cells/well), and after 24 h of incubation, they were treated with yamogenin at the concentrations of 30–100 µg/mL for 5 and 24 h. Oxaliplatin was used as a positive control at a concentration of 20 µg/mL. The experiments were performed in three independent repeats.

The cells were seeded in 12-well plates (1 × 10⁵ cells/well) and treated with the extract at concentrations of 40.0–300.0 µg/mL. After 6 and 24 h of treatment, the apoptotic status of the cells (based on caspase-3 and 7 activation), cellular plasma permeabilization, and cell death were estimated. The cells were stained with a fluorescent reagent that contained a DNA binding dye linked to a DEVD peptide substrate. When active caspases-3 and 7 cleave this complex, the dye is released and binds to DNA. A dead cell marker (7-AAD) was also used in the assay. The cells were analyzed with flow cytometry (Muse Cell Analyzer). The experiments were performed in three independent repeats.
The activity levels of caspases-8 and 9 were determined using a Caspase-Glo 8 or 9 Assay Kit (Promega, Madison, WI, USA) and Glomax Multi+ Detection System (Promega, Madison, WI, USA). The cells were seeded in 96-well plates (1 × 10⁴ cells/well), then after 24 h they were incubated with K. daigremontiana water extract at concentrations of 10–300 µg/mL for 2–24 h. The experiments were performed in three independent repeats.