The apoptotic effects of the fraction (fraction C) on HeLa and SKOV-3 cells were estimated using Annexin V and Dead Cell Assay Kit (Merck Millipore). The cells were seeded in 12-well plates at a density of 1 × 105 cells/well and treated with the fraction at concentrations of 3.0–100.0 µg/mL. The fraction solvent – methanol was added to the control cells at a concentration of 1.0% (v/v). After treating, the cells were prepared as described previously (Stefanowicz-Hajduk et al. 2016 (link)) and analyzed by flow cytometry with Muse Cell Analyzer (Merck Millipore), following the protocol provided by the manufacturer. The experiments were performed in three independent repeats.
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