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Fig. 2. The p38 MAPK pathway is involved in the early expression of Nkx2.5. (A) One of two blastomeres (upper panel) or two of two blastomeres (lower panel) were injected with p38α AMO, and the expression of Nkx2.5 and GATA4 was detected by in situ hybridization of stage 13 (top) and stage 33 (bottom) embryos. (B) p38α AMO was injected to embryos, and gene expression was assessed in three cardiac explant assays; DMZ explants (left), AC explants from Alk4-expressing embryos (middle) and VMZ explants from LiCl-treated embryos (right). At stage 15, total RNA was isolated and RT-PCR analysis was performed. (C) Transcripts encoding inactive MKK6 (MKK6A) were injected to one cell embryos, and DMZ explants (n = 18) were isolated and were grown to stage 15. Total RNA was isolated and RT-PCR analysis was performed. (D) Upper panel: Transcripts encoding MKK6E were injected to one of two blastomere embryos (n = 24). Embryos were allowed to develop to stage 14 and in situ hybridization was performed with a probe to Nkx2.5. The injected side was recognized by the staining of a β-Gal lineage tracer. Lower panel: Transcripts encoding MKK6E were injected to one cell embryos, VMZ explants (n = 18) were isolated and were grown to stage 14. Total RNA was isolated and RT-PCR analysis was performed. |
