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. 2018 Sep 28;6:126. doi: 10.3389/fcell.2018.00126

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Copyright © 2018 Sun, Chen, Zhang, Munisha, Dougan and Sun.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Mga binds to Bs69 and disrupts the Bs69–Bmpr1a interaction. (A) Increasing dose of pCS2-Mga (1, 2, 3 μg) along with 1 μg pCS2-HA-Bs69, 1 μg pCS2-Bmpr1a-FLAG were transiently co-transfected into 293T cells, and co-immunoprecipitation of HA-Bs69 and Bmpr1a-FLAG was analyzed. (B) Increasing dose of pCS2-Bmpr1a-FLAG (1, 2, 3 μg) along with 1 μg pCS2-HA-Bs69, 1 μg pCS2-Mga were transiently co-transfected into 293T cells, and co-immunoprecipitation of HA-Bs69 and Mga was analyzed. (C) One-cell-stage wild-type embryos injected with 50 pg mRNA encoding HA-Bs69 or mixture of mRNAs encoding both HA-Bs69 and mouse MGA, or co-injected with 50 pg mRNA encoding HA-Bs69 with 100ng pCS2-FLAG-Mga or pCS2-FLAG-MgaΔ. Immunoblot analysis of pSmad1/5 levels was performed with lysates from the injected embryos at 8 hpf. (D) Relative quantification of pSmad1/5 levels of panel C. (E) One-cell-stage wild-type embryos injected with 50 pg mRNA encoding HA-Bs69 or mixture of mRNAs encoding both HA-Bs69 and mouse or zebrafish MGAs, and the ventral tail fin phenotypes of these embryos at 48 hpf were shown.