DEV02674 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. wg expression in srt^7E4-mutant tissues. (A) Wg expression as measured by wg-lacZ in blue. (B) GFP (green) marks srt^7E4/+ heterozygous cells that are outlined in both A and B. Notice the expansion in the number of wg-expressing cells in srt^7E4-mutant tissue. Although an expansion in the number of wg-expressing cells at the dorsoventral boundary is observed, the level of wg expression in these cells is diminished. Hence, the accumulation of Wg protein within srt^7E4-mutant cells cannot be accounted for by an increase in wg transcription.

• Supplemental Figure 2 -

  Fig. S2. Expression of Wg, Sen and Distal-less in wing discs of srt P-element excision alleles. (A-O) Wg staining in green (A,D,G,J,M), Sen (a short-range target of Wg signaling) in blue (B,E,H,K,N) and Distal-less (Dll, a long-range target of Wg signaling) in red (C,F,I,L,O). All alleles of srt balanced over TM6B, Tb were crossed to Df(3L)ED4470 (68A6;68E1)/TM6B, Tb. Wing discs from non-Tb larvae were dissected and stained as described. The transheterozygous combinations are (A-C) wild type, (D-F) srt^11001-1/Df(3L)ED4470, (G-H) srt^11001-2/Df(3L)ED4470, (J-L) srt^01593-1/Df(3L)ED4470 and (M-O) srt^7E4/Df(3L)ED4470. In srt^11001-2/Df(3L)ED4470 wing discs (H) approximately half showed some Sen expression at the dorsoventral boundary, which corresponded to a wild-type pattern of Wg expression. Expression of the Wg long-range target Dll yielded considerably more variability, even within a given transheterozygous combination. However, we consistently observed that Dll expression was reduced and occasionally was almost-completely absent. The reason for this variability is probably due to the perdurance of maternally contributed Srt and the lower levels of Wg protein required to activate Dll expression in target receiving cells. In all crosses, homozygous pupae were present, confirming that, zygotically, srt is pupal lethal. The panels for wild type, srt^11001-1 and srt^11001-2 were obtained from staining the same wing disc with mouse anti-Wg, guinea pig anti-Sen and rabbit anti-Dll (1:400, G. Boekhoff-Falk). For srt^01593-1 and srt^7E4, Wg and Sen panels were from the same wing disc and Dll staining was performed with an independent wing disc of the same genotype using mouse anti-Dll (1:1000, I. Duncan).