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Fig. 2. Sox9 is required for Snail2 induction downstream of
BMP signal. (A) BMP4 treatment (lower panels) of the neural plate
explants effectively induces Sox9 expression and cell dispersal. (B)
Neural plate explants were co-transfected with Snail2 and
GFP expression vectors, and stained with anti-Sox9 antibody (upper
panels) or phalloidin (lower panels). Snail2 misexpression fails to
induce either Sox9 protein expression or epithelial-mesenchymal transition
(EMT). (C) Sox9 transfection strongly induces Snail2
expression in neural plate explants and subsequent EMT indicated by an
extensive cell dispersal, in the absence of BMP4. Insets indicate higher
magnification, showing typical morphology of Sox9-transfected cells.
(D) Electroporation of GFP and Sox9-En-nuc expression
constructs was performed on quail embryos at stage 5, and the embryos were
cultured for 7 hours. Misexpression of Sox9-En-nuc expression vector
reduces the Snail2 mRNA expression (arrowheads), compared with the
untransfected side, or a control embryo transfected with GFP alone.
(E) Sox9 is required for Snail2 induction by BMP4. Neural
plate explants, transfected with double-stranded RNA corresponding to
Sox9 sequence (Sox9 dsRNA), show little expression of Snail2
or cell dispersal, in the presence of BMP4. Transfection of mutated dsRNA
(Sox9mut dsRNA) has no effect (Sox9+Sox9 dsRNA) and the
effect of Sox9 dsRNA is cancelled by a co-transfection of
Sox9 (Sox10+Sox9 dsRNA), while co-transfection of
Sox10 fails to cancel the effect of Sox9 dsRNA.
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