HCmel12 cells were seeded into six-well plates (1 × 105 cells per well) in complete RPMI medium. Six hours later, FR was added at various concentrations. Control cells received vehicle only. After 72 hours, cells were collected and analyzed for apoptosis induction and cell cycle arrest using standard protocols with annexin V (BD PharMingen) and PI. Data were acquired with a FACSCanto flow cytometer (BD Biosciences) and analyzed with FlowJo software (Tree Star, V7.6.5 for Windows).

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