dmrt2b maintains BMP signaling in the pharyngeal region by inhibiting cv2 expression. (A) Expression of BRE-driven EGFP in the pharyngeal region of Tg(BRE:EGFP;sox10:mCherry-CAAX) transgenic embryos injected with 4 ng cMO or dmrt2b MO at 36 hpf. (B,C) p-Smad1/5/8 levels were decreased in the pharyngeal region of dmrt2b mutants (B) and morphants (C) at 36 hpf. Embryos were stained with the indicated antibodies. Nuclei were counterstained with DAPI (blue). (D,E) The expression of cv2 in dmrt2b morphants were examined by in situ hybridization (D) and qRT-PCR (E). All data are presented as the mean of three independent experiments. Error bars represent s.d. Significance was analyzed using unpaired t-tests. **P<0.01. (F) Co-injection of 100 pg cv2 MO with 4 ng dmrt2b MO partially rescued the EGFP expression in Tg(BRE:EGFP;sox10:mCherry-CAAX) transgenic embryos. (G) p-Smad1/5/8 levels were partially rescued in the pharyngeal region of dmrt2b mutants co-injected with 100 pg cv2 MO at 36 hpf. Embryos were stained with the indicated antibodies. Nuclei were counterstained with DAPI (blue). ov, otic vesicle; p-S1/5/8, phosphorylated Smad1/5/8. Scale bars: 50 µm (A–C,F), 20 µm (G).
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