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Figure 1


Fig. 1. XMAN1 inhibits the activin/nodal pathway in the Xenopus embryo. (A) Whole-mount in situ hybridization for chordin (chd) at stage 10.5. MO, antisense morpholino oligonucleotides against XMAN1; 4mmMO, four-base-mismatched MO. XMAN1 mRNA (1 ng/embryo) or MO (75 ng/embryo) was injected into two dorsal blastomeres at the four-cell stage. Arrowheads indicate the dorsal lip. (B) Suppression of Xnr1-dependent mesodermal induction by XMAN1. RT-PCR of animal caps for chd and goosecoid (gsc), dorsal mesoderm markers and Xenopus brachyury (Xbra), a pan-mesodermal marker, at the equivalent of stage 10.5. Xnr1 mRNA (100 pg/embryo) and either mRNA for XMAN1 (2 ng/embryo), C-terminal (CT)-XMAN1 (500 pg/embryo, equivalent amount in mol to XMAN1) or CT-truncated ({Delta}CT)-XMAN1 (1500 pg/embryo, equivalent amount in mol to XMAN1) were injected. FGFR is the loading control. (C) Suppression of activin/nodal-dependent Xnr1 intron1-luciferase (Int1) activation by XMAN1 constructs. A control of GL3-luciferase or an Int1 construct (100 pg/embryo) was injected together with a Renilla luciferase plasmid (2 pg/embryo), Xnr1 mRNA (100 pg/embryo) and mRNA for XMAN1 (1 ng/embryo), CT (250 pg/embryo) or {Delta}CT (750 pg/embryo).