nradd is transcriptionally regulated by Wnt/β-catenin signaling during development. (A) Whole-mount in situ hybridization (WMISH) shows that nradd expression is downregulated in transgenic embryos expressing Axin1 (hs:Axin1) and upregulated in transgenic embryos expressing Wnt8a (hs:Wnt8a) at gastrula (80% epiboly), somitogenesis (10-somite) and organogenesis (22 hpf) stages. (n (80% epiboly): wild type 21/21 embryos, hs:Axin1 18/20 embryos, hs:Wnt8a 16/17; n (10-somite st.): wild type 24/24 embryos, hs:Axin1 24/24 embryos, hs:Wnt8a 28/30; n (22 hpf st.): wild type 19/19 embryos, hs:Axin1 23/25 embryos, hs:Wnt8a 19/20) hs: heat shock, fix: fixation, st: stage, hpf: hours post-fertilization. Three independent experiments were conducted. (B–D) nradd and sp5l expression levels determined by qPCR in hs:Axin1 and hs:Wnt8a transgenic embryos are shown relative to those in wild-type embryos at (B) gastrula, (C) somitogenesis and (D) organogenesis stages. Statistical significance was evaluated using an unpaired t-test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Error bars represent ± standard deviation (SD, n = 3). Three independent experiments were conducted.
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