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Fig. 5. Efnb2 loss-of-function mutation elicits enlarged DA and
underdeveloped CV, resembling Notch gain-of-function mutant
morphology. (A,B) Whole-mount CD31 staining of E9.0 (17 ss)
embryos. Arrows, DA; arrowheads, CV. (C,D) Higher magnifications
of A, B, respectively. Note the enlarged DA (arrows and white brackets) and
underdeveloped CV (arrowheads and blue brackets) in the
Efnb2-deficient embryo (B,D). (E,F) 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 (15-17 ss) embryos. A total of 2122 and 1714 ECs were counted in control
and mutant embryos, respectively. Total EC number between mutants and controls
is decreased (n=3, P=0.02). The proportion of ECs in DA (da,
red) over primordial ACVs (p-acv, blue) is significantly increased
(n=3; *P=0.02) in mutants (F), when compared with
controls (E). (G,H) CD31 (red) and EphB4 (green) staining of
cross-sections of E8.75 (15 ss) embryos. EphB4+ ECs are present in
the DA (arrow) of the enlarged Efnb2-deficient DA (H).
(I,J) Whole-mount CD31 staining shows enlarged DA and reduced CV
in E8.75 (16 ss) embryos with Tie1-cre-mediated deletion of Efnb2
(J). (K,L) Higher magnifications of I, J, respectively. Arrows
and white brackets, DA; Arrowheads and blue brackets, ACV. Scale bars: 200
µm.
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