Fig. 2. Organizer genes are expressed but some neural markers are reduced in Xnr3^– embryos. (A) Organizer gene expression was analyzed at gastrula and early neurula stages (stage 10, 10.5, 11, 12, 14). Xnr3 MO does not affect the expression level of organizer genes. (B) Neural markers were analyzed at stage 28 NCAM and en2 expression was repressed in MO-injected embryos. 10 ng (for late markers), or 20 ng (for both early and late markers) was injected into the dorsal marginal two cells at the 4-cell stage. In each case, ornithine decarboxylase (ODC) was used as a loading control (data not shown), and expression of each gene was normalized to the level of ODC expression. (C) chd and gsc are not expressed in the correct region in MO-injected embryos. Expression pattern of chd (top two rows), and gsc (bottom two rows) in uninjected (upper row) and MO-injected (lower row) embryos. chd was expressed normally in MO-injected embryos at stage 11, and continued to be expressed around the dorsal rim of the blastopore even at stage 14. Gsc expression in MO-injected embryos also remained adjacent to the blastopore at stage 14. 20 ng of MO was injected into the two dorsal animal cells of stage 8 albino embryos, and the expression pattern was analyzed by whole-mount in situ hybridization.

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