Fig. 9. Gene expression in the embryos injected with morpholino-modified (MO) antisense oligonucleotides. Injected MO is shown in the upper right and the probe used is indicated at the bottom. Lateral views of the head region at 26 hpf. Embryos were injected with 4mis-MO (A,F,I,K), fgf8-MO (B,G), fgf3-MO (C,H,J,L), fgf8-MO and fgf3-MO (D), and fgf3-MO and fgf3 mRNA (E). (A-E) Wild-type embryos were used. Injection of either fgf8-MO (B) or fgf3-MO (C) reduces nk2.1b expression. However, reduction of nk2.1b is further enhanced when both fgf8-MO and fgf3-MO was co-injected (D). (A'-C') en2 expression after 4mis-MO (A'), fgf8-MO (B') and fgf3-MO (C') injection. en2 expression is reduced in fgf8-MO-injected embryo but remains unchanged in fg3-MO-injected embryo. (E) nk2.1b expression is rescued by co-injection of fgf3-MO and fgf3 RNAs. (F-H) Embryos from ace heterozygous parents were used; fgf3-MO but not fgf8-MO injection enhances the reduction of nk2.1b in some embryos (17/58). (I-L) Injected embryos from ace heterozygous parents were double-stained with en2 and emx1 (I,J) or dlx2 (K,L), and homozygous mutants were identified by a lack of en2 expression in the MHB. An arrowhead in I indicates the subpallial telencephalon, which is negative for emx1. Dots indicate the boundary between the telencephalon and ventral diencephalon. Scale bar: 50 µm. (M) Western blot analyses of translation of injected fgf3-myc. RNA injections were carried out at the one-cell stage. The homogenates were prepared from injected blastula. Anti-Myc antibody detects a major band about 50 kDa which is not present in control injection (GFP RNAs). Relative intensity of the major band is shown at the bottom of each lane. Relative intensity was normalised by a nonspecific minor band that appears in all lanes and insensitive to morpholino injection. (N) Morpholino oligonucleotides used in the present study. Red letters in 4mis-MO indicate mismatch nucleotides to fgf3-MO.