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Expression of wnt5a/5b and wnt11/11r. (A-L) Whole-mount in situ hybridization. Lateral views at 1-cell stage (A,D,G,J), animal pole views with dorsal to the right at 60% epiboly (B,E,H,K), and lateral views with anterior to the left (C,F,I,L). The detection of wnt5b (Rauch et al., 1997), wnt11 (Heisenberg et al., 2000) and wnt11r (Matsui et al., 2005) was described previously. The wnt5a probe was prepared by RT-PCR. (M-P) Quantification by qRT-PCR. Expression levels of wnt5a/5b and wnt11 at 60% epiboly, and wnt11r at 26 hpf, were taken as 100. Primers: wnt5a, 5′-GAACTGCAGCACCGTCGACAACTC-3′ and 5′-GAACTGCAGCACCGTCGACAACTC-3′; wnt5b, 5′-GCCTATCTCAGGGTCAGAGGAA-3′ and 5′-CGCCCCCTCTCCAATATAAAC-3′; wnt11, 5′-CACCCCTGACCTGGCCAAAGGGAC-3′ and 5′-GGAGATGGTGCTGATGTCTTGA-3′; wnt11r, 5′-TCTCACGCCAACAAGCTCAT-3′ and 5′-CCAGAAACACCGTGGCATT-3′. Scale bars: 200 μm.
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