|ZFIN ID: ZDB-IMAGE-140422-39|
sfrp1a/sfrp5 positively regulate Wnt signaling. To determine the function of Sfrp proteins in regulating Wnt signaling, we examined wnt ligand mRNA expression and a transgene reporter of Wnt signaling using whole mount in situ hybridization and quantitative PCR. In situ hybridization for eGFP mRNA in the Tg(TOP:dGFP) line shows a decrease in eGFP expression in sfrp1a/5 MO co-injected embryos, indicating a decrease in Wnt signaling at 28 hpf (A,B). The effects of Sfrp depletion on eGFP expression were confirmed using quantitative RT-PCR (C, NNN, p<0.001). In situ hybridization of Wnt ligands wnt2 and wnt8b at 28 hpf show an expansion in the expression domain of wnt2 and wnt8b in the dorsal retina in sfrp1a/5 MO injected embryos (D–G). qRT-PCR confirmed a 1.33 fold change in wnt2 and 1.38 fold change in wnt8b expression (H, NNN, p<0.005).
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