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Figure 5: Pitx2 expression is affected by HCN4-DN and ZD7288.
(A) Representative images of approximately stage 28 embryos assayed for
Pitx2 expression by in situ hybridization. Left orientation of the embryo is indicated at
the bottom of the image. Red line indicates the anterior-posterior spread of the Pitx2
expression and yellow dotted line indicates the area of the Pitx2 expression (i) is no
probe (negative) untreated control (ii) Control embryos with Pitx2 signal – yellow
arrow. (iii) embryos injected with HCN4-DN mRNA in both blastomeres at 2-cell stage
with Pitx2 expression - yellow arrow, (iv) ZD7288 treated (100μM stage1-10) embryo
with Pitx2 expression - yellow arrow. Scale bar = 0.25 mm. (B) Quantification of
anterior-posterior spread of Pitx2 expression (as indicated by red lines in A) in embryos showed a significant reduction in the spread of Pitx2 expression in HCN4-
DN mRNA injected and ZD7288 treated embryos. N=20, Data was analyzed by oneway
ANOVA. ***p<0.001. (C) Quantification of area of Pitx2 expression (as indicated
by yellow dotted lines in A) in embryos showed a significant reduction in the area of
Pitx2 expression in HCN4-DN mRNA injected and ZD7288 treated embryos. N>25
Data was analyzed by one-way ANOVA. ***p<0.001, **p<0.01. |
