Cluvenone increases total Cyt c levels and induces cytosolic release in CEM cells. Cells were treated with 1 μM cluvenone or 0.1% DMSO for 7 h, followed by subcellular fractionation using a digitonin-based extraction method. Cytochrome c in cell extracts was measured by an ELISA assay with absorbance measured at 405 nm with correction at 580 nm. All conditions were performed in duplicate. Levels of Cyt c in samples was determined by comparison to standards and all values were reported per 10 × 106 cells. Cyto −, cytosolic extract from untreated cells; Mito −, mitochondrial extract from untreated cells; Cyto +, cytosolic extract from cells treated with cluvenone; Mito +, mitochondrial extract from cells treated with cluvenone.