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Fig. 3 Transient transgenic analysis of zic2a-zic5 genomic regions. (A) Conserved DNA regions (thick lines) at the zic2a-zic5 locus. P2 and P5, each approximately 300 nt, are 70% identical with Fugu. IG (840 nt) is 49% identical to Fugu, and D5 (535 nt) is 60% identical to Fugu. Eight consensus Tcf/Lef-binding sites were found in IG and six in D5 (asterisks). (B) Transgenic constructs tested in transient assays. (C,D) Dorsal midbrain and hindbrain expression of Gfp in representative zic5D5:gfp and zic2aD5:gfp transient transgenics. Insets are dorsal views of the embryos shown. Asterisks mark mid-hindbrain organizer. (E-G) gfp RNA in representative transient transgenics. In embryos injected with 25 pg of zic2aD5:gfp (E) or zic2aD5ΔIG:gfp (F), expression was concentrated in dorsal midbrain and hindbrain. Embryos injected with 25, 50 or 70pg of zic2a:gfp (G) did not show enhanced expression. All views are lateral, except for insets.