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Fig. 4. Notch1 loss-of-function mutation elicits smaller DA and
enlarged CV regions. (A,B) CD31 (red) and EphB4 (green)
staining of cross-sections of E9.0 (15 ss) embryos shows remnant DA (arrows)
and expanded primordial ACV areas (arrowheads) in the mutant (B). Note the
increase in EphB4-expressing ECs in the primordial ACVs and DA areas (B).
(C,D) Quantitative analysis of EC distribution. Total ECs,
including those in the DA, CV and capillaries, were counted from the
cross-sections of the anterior region of E8.75 (12-15 ss) embryos. A total of
1336 and 1385 ECs were counted in control and mutant embryos, respectively.
Total EC number between mutants and controls is comparable (n=4,
P=0.52). The proportion of ECs in the DA (da, red) over primordial
ACVs including capillaries (p-acv, blue) is significantly decreased
(n=4; **, P=0.0007) in the mutants (D), compared
with the controls (C). (E,F) Whole-mount CD31 staining shows
reduced DA (arrows) and enlarged CV (arrowheads) in E8.75 (13 ss) embryos with
Tie1-cre-mediated deletion of Notch1. (G,H) Higher
magnifications of E, F, respectively. Arrows and white brackets, DA;
arrowheads and blue brackets, ACV. Scale bars: 200 µm.
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