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Fig. 6. Nkx3.2 represses Runx2 expression. (A) Nkx3.2 and Runx2 are
expressed in a reciprocal pattern in the developing cartilage elements of the
limb. ISH for the indicated probes is displayed for E8 chick limbs. In the
cartilage element, Runx2 expression is most intense where Nkx3.2 expression is
diminished (marked by arrowheads in a,b). (B) Forced expression of
Nkx3.2 represses Runx2 expression in vivo. The right wing buds of E3 chick
embryos were infected with either RCAS(A)-Nkx3.2 (b,d) or
RCAS(A)-Nkx3.2-
C-VP16 (f,h) viruses. Embryos were harvested at either
E8.5 (a-d) or E8 (e-h), and sections of the infected or contralateral control
wings were hybridized with the indicated probes. (C) The intensity of
the Runx2 ISH signal displayed in B and in two other experiments was
quantified, in both the peripheral (blue rectangles in B) and central regions
(orange rectangles in B) of the cartilage in both control and infected wings,
as indicated. (D) Forced expression of Nkx3.2 represses Runx2
expression in presomitic mesoderm explants in vitro. Explants of presomitic
mesoderm were either cultured for 2 days in either Shh (lanes 1 and 3) or
infected with RCAS-Nkx3.2 (lanes 2 and 4). The explants were cultured in Bmp4
for either an additional 6 (lanes 1 and 2) or 12 (lanes 3 and 4) days.
Expression of indicated markers was assayed by RT-PCR.
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