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Fig. 4. Molecular genetic and vital dye lineage marking of serosal EMT. (A-F)
Localization of Wt1 (green) and ß-Gal (red) proteins during gut
development. Wt1 expression is largely confined to the serosal mesothelium in
the embryo (A-C) and newborn (NB0; D-F). The anti-ß-Gal antibody marks
cells in the subserosal space of the developing gut tube (arrows in B,C,E,F).
Both Wt1 and ß-Gal are expressed in most cells of the serosal mesothelium
(C,F, arrowheads). (G-K) CCSFE (green) was used to vitally label surface cells
of gut explants from E12.5 Wt1-Cre; Rosa26R embryos. (G) At day 0 of culture,
surface cells were positive for CCFSE (area framed by arrowheads), but the
subjacent mesenchyme is negative. (H,I) After two days of culture,
CCFSE-positive cells (arrowheads) at the gut surface co-label with Wt1.
CCFSE-marked cells in the subserosal space are not positive for anti-Wt1
antibody (arrows). Also, many mesenchymal cells label with both
anti-ß-Gal antibody and CCFSE (arrows, J,K). ß-Gal-positive cells
unmarked by CCFSE are also seen in the mesenchyme (open arrowheads). Note a
small number of CCFSE-labeled cells without the ß-Gal marker (filled
arrowheads). Scale bars: 20 µm.
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