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. 2008 Dec 29;18(8):1155–1166. doi: 10.1089/scd.2008.0391

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Copyright 2009, Mary Ann Liebert, Inc.

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FIG. 3. — Purified side population (SP) cells can be expanded in culture under standard embryonic stem cell (ESC) conditions producing SP and non-SP cells that express markers of undifferentiated ESCs. (A–E) Indirect immunofluorescence microscopy of cultures derived from sorted SP cells. (A) General appearance of culture, as visualized in phase contrast mode. (B) SSEA1 immunostaining. (C) Nuclear staining with DAPI. (D) Merge of SSEA1 and DAPI emissions. (E) Isotype control. Scale bar, 100 μm. (F) Hoechst FACS profile of cells from SP-derived culture. Y-axis: Hoechst red emission; X-axis: Hoechst blue emission. Gate represents SP cells. (G–J) FACS analysis of ESC undifferentiation antigens in the SP fraction of cells shown in F. Y-axis: normalized frequency of events; X-axis: fluorescence intensity for the indicated antigen. Emission peaks for the indicated antigen and Isotype controls and shown.