Skip to main content
. 2006 Jun 1;20(11):1470–1484. doi: 10.1101/gad.1416106

Figure 1.

Figure 1.

Identification of Evf-2 transcripts. (a) The ultraconserved Dlx-5/6 intergenic enhancer ei is transcribed in vertebrates. Human and mouse ESTs were found in the database (sky blue). ei-containing ESTs were not found in the zebrafish and chick databases. The right panel shows that two different primer sets (A and B) detect ei-containing transcripts in cDNAs made from zebrafish embryos or chick embryonic brains. (b) Genomic organization of the mouse Evf gene. Evf-1 and Evf-2 result from alternate transcription initiation, alternative splicing of exon 3, and alternative polyadenylation. Exon 2, included in Evf-2 but absent from Evf-1, is partly encoded by the conserved intergenic enhancer ei (red). (c) Schematic representation of Evf-1 and Evf-2. Common regions are shown in blue. Evf-2 nucleotides 116–459 are encoded by the conserved intergenic enhancer, ei (red). Antisense RNA probes (shown as pink, blue, and red dashed lines) spanning different regions of Evf-1 and Evf-2 were used for Northern, RNase protection, and in situ hybridizations, as indicated. (d) Northern analysis of total and polyadenylated RNAs isolated from rat E13.5 embryonic telencephalon, using probes generated against Evf-1 and Evf-2 (as indicated in c). (D) Dorsal; (V) ventral. The full-length Evf-1 probe detects the 28S rRNA in total RNAs from both dorsal and ventral RNAs, showing that the absence of Evfs in D RNAs is not due to degradation. (e) RNase protection. A 450-nt probe spanning the Evf 2/common region junction (blue) detects RNase-resistant bands in ventral (V) but not in dorsal (D) rat E13.5 embryonic telencephalic RNAs or tRNA. (f, panels i,ii) Section in situ hybridization of tissues from rat ventral forebrain at E11.5 using the 5′specific Evf-2 probe (pink). (Panel iii) Dense Evf-2 staining colocalizes with the nuclear stain DAPI. (g) Section in situ hybridization of mouse ventral forebrain at E12.5 using the following antisense probes: Dlx-5 (panel i), Evf (1 + common region) (panel ii), Shh (panel iii), Dlx-6 (panel iv), Evf-2 (panel v), and Dlx-2 (panel vi). (VZ) Ventricular zone; (LGE) lateral ganglionic eminence; (MGE) medial ganglionic eminence; (D) dorsal; (V) ventral. The full-length rat Evf-1 cDNA sequence is available on GenBank (accession no. AY518691).