Suv4-20h double-null ES cells have elevated Oct4 and lower CXCR4 protein levels before and during differentiation.

<p>Wild-type and Suv4-20h DKO ES cells were grown undifferentiated in LIF-containing medium, or differentiated <i>in vitro</i> 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 µm). 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 2×10⁴ 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.