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Fig. 3. (A) Flow cytometric analysis of PI-stained Kit^+/+ and Kit^W-lacZ^/^W-FKB cells at different times after LIF withdrawal. The increase in the subG-phase fraction and decrease in G0-G1 and G2-phase fraction was observed in Kit^W-lacZ/W-FKB cells 3 days after induction of differentiation. (B) Ethidium-bromide-stained genomic DNA isolated from undifferentiated (d0) Kit^+/+ (+) and Kit^W-lacZ/W-FKB (-) ES cells and 1, 2, 2.5, 3 and 3.5 days after LIF withdrawal. (C) AnnexinV (x axis) and PI (y axis) staining of Kit^+/+ and Kit^W-lacZ/W-FKB ES cells grown in the presence of LIF. The numbers shown in each quadrant represent the percentage of cells staining for Annexin only (red), PI only (green) and both Annexin and PI (magenta). (D) Western blot analysis of protein lysates isolated from undifferentiated (d0) Kit^+/+ (+) and Kit^W-lacZ/W-FKB (-) ES cells and four days (d4) after LIF withdrawal. Blots were sequentially probed with an antibody specific to BCL2 protein then an anti-GAPDH antibody as a loading control.

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