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Fig. 6. Variability of Cdx2, but not Nanog, may be generated by asymmetric cell
divisions at the 8-cell stage. (A,B) Cdx2 and Nanog
patterning of blastomeres isolated at the 8-cell stage and analyzed at 70 (A)
and 78 (B) hp-hCG. (C) Scatter plot illustrating the normalized mean
fluorescence intensity of Cdx2 (left panels) and Nanog (right panels) in
sister blastomeres at 70 (C1) and 78 (C2) hp-hCG. The `smaller' (at 70 hp-hCG)
or `inside' (at 78 hp-hCG) cell is plotted on the x-axis, the
`bigger' or `outside' one on the y-axis. A filled circle represents a
doublet with clear asymmetry; an empty circle represents a doublet with slight
asymmetry; an empty rhombus represents symmetric doublets. Each symbol
represents a 2/16 doublet; different colors represent different embryos. Red
lines mark the background fluorescence level. (D) Cdx2 and Nanog
patterning at 90 hp-hCG in blastomeres isolated at the 8-cell stage.
(E) Cdx2 and Nanog patterning of blastomeres isolated at 8-cell and
re-isolated at the 16-cell stage. A cross marks an apolar cell; black
triangle, a polar cell; asterisk, eccentric localization of the nucleus;
arrowhead, the pole of intense actin staining in polar cells. DNA staining
provides a reference value. Actin (red) marks the cell membranes. All images
are single optical sections. Scale bar: 20 µm. (F) Scatter plot
illustrating the Cdx2 variance in relation to Nanog levels of polar (filled
triangles) and apolar (crosses) cells. Each symbol represents one blastomere.
Black lines and dotted lines represent the average protein levels of polar
cells and apolar cells, respectively. Error bars indicate s.d.
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