Fig. 3. eyc encodes a Drosophila p47 homolog and eyc is misexpressed in early pupae. (A) A Northern blot shows a 2 kb genomic probe detects a 1.1 kb mRNA, the levels of which are elevated at 30% eyc and P1363/Df(2R) Px2 (P/Df) flies relative to wild type (CS); RP49 was used as loading control. (B) A Western blot shows the anti-Eyc antiserum detects Eyc protein, which is more abundant in 30% p.d. eyc¹ retinal extract compared with wild type at the same developmental stage; ß-Tubulin was used as loading control. (C) Molecular map of eyc locus. 60D1 represents genome project predicted genomic map of eyc locus; predicted genes nearby eyc are denoted as white bars with pointed ends indicating transcript orientation. The eyc locus is shown above, with the white bar indicating the ORF. The eyc allele has two 3' point mutations (arrowheads). The P-element insertion site of eyc^P allele is shown. The eyc null allele eyc^l39 is shown below 60D1; brackets mark the determined deletion and broken lines indicate undetermined potential deletion. The DNA fragment used to construct the pUAS-eyc transgene is shown as a hatched bar. (D) ClustalW alignment of the predicted Eyc amino acid sequence with two related proteins, rat p47 (human p47 sequence is 95% identical) and yeast SHP1. Residues identical in all three species are shown on a red background and are indicated by ‘!’; residues identical or with conserved substitutions in two species are shown on black or blue, respectively, and are indicated by an asterisk. The p47/Eyc family shares about 30% protein sequence identity and 45% similarity. eyes closed sequence data are available from GenBank/EMBL/DDBJ under Accession Number AF170565.