Fig. 2

Induction of caspase-independent apoptosis in FrA-treated cells. a Western blotting analysis of protein extracts of RT112 cells treated with FrA for 48 h. b Cell cycle analysis of RT112 cells treated with FrA for 48 h. Apoptotic cells are detectable as a sub-G1 population. Cell cycle phase distribution was quantified using the Cell Quest Pro software. c, d Flow cytometry analysis of RT112 cells treated with FrA using an annexin-V-FITC/PI double staining (c) and quantification of positive cells (d). Cells were pretreated with 100 μM of the pan-caspase inhibitor zVAD for 1 h and then treated with indicated concentrations of FrA or with anisomycin (Aniso, positive control) for 48 h. Apoptotic cells appearing in the right lower and upper quadrants were quantified using the Cell Quest Pro software