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Figure 8. Suv4-20h double-null ES cells have elevated Oct4 and lower CXCR4 protein levels before and during differentiation. Wild-type and Suv4-20h DKO ES cells were grown undifferentiated in LIF-containing medium, or differentiated in vitro by embryoid body formation. Wt26 - isogenic wild-type ES cell line; B4-2, B7-1 independent Suv4-20h DKO ES cell lines. (A) Morphological appearance (scale bar 100 ). Top row: undifferentiated ES cells (day 0); middle: embryoid bodies at day 6; bottom: cells from embryoid bodies, replated for 24h. (B) Before (day 0) and during (day 6) differentiation, cell lines were stained for Oct4 protein and 204 cells from each condition were subjected to FACS analysis. Red graph: fluorescence of non-specific isotype control; black and green graphs represent the Oct4 protein levels in wild-type and Suv4-20h DKO ES cell lines, respectively. (C) Suv4-20h DKO cells have higher Oct4 protein levels compared to wild-type ES cells and maintain these during differentiation. Median fluorescence intensity was calculated from data in panel (B), error bars indicate SEM. (D) Suv4-20h DKO cells show a reduction in the percentage of CXCR4 + cells at differentiation day 6. Data represent normalized values of percentage of CXCR4+ cells as means of three independent experiments, error bars indicate SEM. doi:10.1371/journal.pgen.1003188.g008 Image published in: Nicetto D et al. (2013) Image reproduced on Xenbase with permission of the publisher and the copyright holder. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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