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Fig. 4. FGF induces Nkx2.5 gene expression without affecting GATA4 gene expression. (A) Expression of FGF3 was detected on stage 13 embryos by whole-mount in situ hybridization (n = 20) (left). Stained embryos were transversely sectioned at the stained plane. One section is presented (right). Anterior transverse sections in stage 13 embryos followed by in situ staining with a probe to Nkx2.5. Nkx2.5 is expressed in lateral mesoderm of anterior section (bottom left). Adjacent sections immunostained with antibodies to phospho-CREB (S133). Red dots represent nuclear phospho-CREB staining. The embryo scheme indicates the plane section. (B) Transcripts encoding truncated FGF receptor (DN FGFR) were injected to embryos and DMZ explants (n = 36) were isolated and grown to stages 14 and 27. Total RNA was extracted and RT-PCR analysis was performed. (C) Transcripts encoding DN FGFR were injected to one cell embryos and the expression of GATA4 and Nkx2.5 was analyzed by in situ hybridization of stage 16 embryos. (D) VMZ explants were dissected from embryos injected with transcripts encoding DN FGFR or from control embryos. At stage 11, basic FGF (150 ng/ml) was added and explants were grown to stage 16. Total RNA was extracted and RT-PCR analysis was performed. |
