GANT61 induces in vitro cell death in ES cell lines. (A) three ES cell lines (TC71, A673 and SKES1) were treated or not for 48 h with GANT61, as indicated. Cell viability was evaluated as described in the material and methods section. For each cell line, the graph indicates the relative cell viability compared to untreated cells. The mean ± SD of at least five independent experiments, each performed in sextuplicate, is presented (** p < 0.005). (B) upper panels: Representative dot plots of SKES1 cells treated or not with 5 or 15 µM GANT61 for 24 h are shown (representative graphs of three experiments). Lower panels: ES cells were treated or not with 5 or 15 µM GANT61 for 24 h. Bars indicate the means ± SD of the relative number of cells in early- or late-phase apoptosis (3 independent experiments). (C) ES cells were treated with 15 µM GANT61 for 24 h. Relative caspase-3/7 activity was evaluated as described in the material and methods section. Bars indicate the caspase-3/7 activity (mean ± SD) of at least two independent experiments, each performed in triplicate (* p < 0.05). (D) ES cells were treated or not with 5 or 15 µM GANT61, as indicated, for 24 h. After incubation, PARP cleavage levels were detected by Western blot analysis. Representative blots of three experiments are shown.