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Fig. 2. Defects in proliferation and differentiation of hepatic epithelial cells
in Tbx3-/- embryos. (A) Morphology of
Tbx3+/+ wild-type and Tbx3-/- mouse embryos (E12.5).
Besides the abnormal development of Tbx3-/- hind limbs (upper and
middle panels), the Tbx3-/- liver was much smaller than
that of wild type (bottom panel). (B-E) Hematoxylin and Eosin (HE)
staining of liver from wild-type (B,D) and Tbx3-/- (C,E) embryos.
(F-I) BrdU immunofluorescent images of wild-type (F,H) and
Tbx3-/- (G,I) liver cells. (J-Q) When compared with
the wild-type liver (J,L,N,P), the number of E-cadherin+ epithelial
cells, but not of N-cadherin+ (E-cadherin-) mesenchymal
cells, was significantly smaller in the absence of Tbx3 (M). Within
Tbx3-deficient liver epithelial cells, primitive hepatic cells and
differentiating hepatocytes that expressed Hnf4
and Alb constituted
only a small population (O,Q), whereas the number of CK7+
cholangiocytes was relatively large (Q). Insets denote individual cells as
labeled with DAPI. (R) Gene expression analysis by qPCR for E12.5 liver
from wild-type, Tbx3+/- and Tbx3-/-
embryos. Two mice of each genotype were analyzed separately. All data were
normalized to the values of a wild-type liver and fold differences are shown.
Bar represents mean ±s.d. (n=3). Scale bars: 200 µm in B,C;
100 µm in F-I; 50 µm in D,E,J-Q.
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