Fig. 1. Enhancement of apoptosis in EBs derived from afadin^–/– ES cells. EBs formed from wild-type and afadin^–/– ES cells were fixed, and apoptotic cells were detected using the TUNEL method and immunostaining with the anti-cleaved-caspase-3 mAb. F-actin (red) and nuclei (blue) were counterstained with rhodamine-phalloidin and DAPI, respectively. (A) EBs at 9 days after the withdrawal of leukemia inhibitory factor (LIF). (B) EBs at 11 days after the withdrawal of LIF. (C) EBs at 15 days after the withdrawal of LIF. Scale bars: 50 µm. The results shown in this figure are representative of three independent experiments.