DEV01869 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Dissection of the 3¢ eye enhancer. (A) Serial dissection of the 16.6 kb, 3¢ genomic fragment that displays enhancer activity in the eye, leg, antenna and wing imaginal discs. Initially convenient restriction sites were used to isolate the 1.9 kb eye-lamina enhancer. Overlapping PCR generated subfragments led to the isolation of 3EE^{390 bp} and 3EE^{194 bp}. The 3EE^{850 bp} and 3EE^{659 bp} were generated based on CNCS predictions and are shown for comparison. Although 3EE^{194 bp} was generated prior to the CNCS analysis, it contains two of the six predicted CNCS blocks and both the So-binding sites. Fragments that have eye specific enhancer activity are shown in blue. The restriction sites are: BamHI (B), EcoRI (R), NotI (N), SpeI (S),and XhoI (X). (B-D) A third instar eye disc from a 3EE-HA-dac larva double-labeled with HA (B, red) and Sens (C, green). (D) A merge of the two channels. HA expression in 3EE-HA-dac eye discs is posterior to the earliest Sens expression and overlaps with anterior Dac, but is rapidly downregulated posterior to the furrow, unlike the GFP expression shown in Fig. 1D, strongly supporting our hypothesis that GFP expression posterior to the furrow represents GFP perdurance and not enhancer activity.