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Fig. 4. Localisation of Nanog, Gata4 and GFP in
PdgfraH2B-GFP/+ blastocysts. (A) In a 32-cell
blastocyst, Nanog and GFP show broad, overlapping expression; Gata4 is
undetectable. (B) Salt-and-pepper distribution of Nanog, Gata4 and GFP
in a 64-cell blastocyst. Gata4 is expressed only in a subset of Nanog-negative
and GFP-positive cells. (C) Partial segregation of Nanog-positive and
Gata4-/GFP-positive cells in a 115-cell blastocyst. Gata4 and GFP are
co-expressed in cells mostly localised to the nascent PrE layer. Nanog is
expressed in the EPI layer and no longer colocalises with Gata4 or GFP.
(D) In a 118-cell blastocyst, PrE markers (Gata4 and GFP) are
co-expressed and mutually exclusive from the EPI marker Nanog; cells
expressing PrE and EPI markers are fully restricted to their respective
layers. Each row represents a single optical section of one embryo. bf, bright
field; green, GFP; white, Nanog; red, Gata4 (A-D); blue, Hoechst. Scale bar:
20 µm. (E) Comparison of the number of Gata4- and Nanog-positive
cells versus total cell number. An abrupt decrease in the number of
Nanog-expressing cells at around the 64-cell stage coincides with the
emergence of Gata4-expressing cells. Thereafter, the number of
Nanog-expressing cells appears to increase only slightly, whereas the number
of Gata4-expressing cells increases approximately linearly. (F)
Comparison of the number of Gata4- and GFP-expressing cells with respect to
total cell number.
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