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Fig. 5. Nkx2.2 regulation of Phox2b. (A) Pancreas from a mouse embryo
harvested at E12.5 stained by immunofluorescence for Nkx2.2 (Cy3, red) and
Phox2b (FITC, green). (B) Lineage tracing was performed by crossing
R26R mice with Wnt1-cre mice. Mouse embryos harvested at
E12.5 were stained for β-galactosidase activity with X-gal (blue), and
stained for Nkx2.2 by immunohistochemistry with peroxidase labeling (brown).
(C) Phox2b mRNA levels were determined by real-time RT-PCR
(TaqMan) with RNA isolated from pancreas (black bars) and stomach (white bars)
from E13.5 embryos with the Nkx2.2 genotypes shown. Values for both
pancreas and stomach are expressed separately relative to the levels of each
in Nkx2.2+/+ embryos. The Phox2b mRNA level in
the stomach was not tested for the Nkx2.2+/- embryos. Data
represent the mean values of four independent experiments performed in
triplicate ±s.e.m. (D-G) The pancreas (D,E) and stomach (F,G) of
Nkx2.2+/+ (D,F) and Nkx2.2-/- (E,G)
embryos at E13.5 were stained by immunofluorescence for Phox2b (Cy3, red) and
Nkx2.2 (FITC, green). An increase in Phox2b staining was observed in the
pancreas from Nkx2.2-/- embryos (compare D and E) but not
in the stomach (compare F and G). *P<0.01;
**P<0.001, compared with Nkx2.2+/+
embryos by Student's t-test. Scale bars: 50 µm.
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