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. 2004 Nov 22;167(4):723–734. doi: 10.1083/jcb.200405144

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Copyright © 2004, The Rockefeller University Press

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Figure 2. — Treatment with ceramide analogues eliminates Oct-4(+)/PAR-4(+) cells in EBs derived from mouse or human ES cells. (A) Mouse EBs were incubated for 24 (middle) or 48 h (right) with 80 μM of S18 showing ongoing cell death in the peripheral and central region of the EB. (B) The center of S18-treated mouse EBs shows intensive costaining of Annexin V (FITC, green)–, PAR-4 (Cy3, red)–, and Oct-4 (Cy5, blue)–positive cells. (C and D) S18-treated human EBs were stained for apoptotic cells using TUNEL assays (FITC, green) and confocal immunofluorescence microscopy was used to detect PAR-4 (Cy3, red)– and Oct-4 (Cy5, pink)–positive cells. Arrows show apoptotic Oct-4(+)/PAR-4(+) (arrowhead 2) or Oct-4(−)/PAR-4(+) (arrowhead 1) cells.