Xenbase Image ID: 159185

Fig. 4. Ventralized development from depletion of maternal Rnf146. (A) RT-qPCR results showing that a specific antisense ODN (AS3m) depleted the maternal store of rnf146 mRNA in oocytes in a dose dependent manner, but had limited effects on the expression of rnf146 mRNA in early gastrula stage (stage 10) embryos. (B) Representative embryos at stage 22/23 from a typical host experiment where 6–10 ng Rnf146 AS3m were injected. DAI: dorsal anterior index. Normal embryos are scored as DAI 5; the fully ventralized embryos were scored as DAI 0; partially ventralized embryos were scored DAI ranging from > 0 lesser than 5. (B′) Histographic presentation of DAI scores from maternal Rnf146 depletion experiments representatively shown in (B). ***P < 0.001 by Student t-test. Embryos from three repeated experiments were pooled together for DAI scoring per the scales depicted in (B). Numbers of embryos for each treatment group are shown on the top of each histogram. (C) RT-qPCR examination of organizer gene expression in control and 6–10 ng AS3m injected embryos at stage 10. Data shown as mean + SD. Error bars were obtained through the statistical treatment of technical replicates. (D, D′) Assessment of specificity of maternal Rnf146 depletion using the antisense ODN AS3m. (D) Representative embryos at the early tadpole stage with indicated treatments. 200 pg mRNA encoding Rnf146 was injected into AS3m (8 ng)-injected oocytes before progesterone treatment. 200 pg mRNA b-catenin was injected into subequatorial zone of maternal Rnf146-depleted embryos at the 2-cell stage. (D′) Histographic presentation of DAI scores from maternal Rnf146 depletion experiments representatively shown in (D). ***P < 0.001 by Student t-test. Image published in: Zhu X et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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