DEV02737 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Expression Patterns. (A) Wild-type adult wing and haltere. (B) dpp-lacZ expression is weaker and narrower in the haltere when compared with the wing. Mad-P staining shows two peaks in the wing, but only one in the haltere (Crickmore and Mann, 2006; de Navas et al., 2006). (C) Intensity traces of Mad-P and dpp-lacZ from the haltere in B. dpp-lacZ expression is graded in the haltere, but the signal decays more rapidly than the Mad-P signal in the A compartment. The horizontal blue dotted line marks the low point of Mad-P staining. The vertical black dotted line marks the depletion of the dpp-lacZ signal, a point at which Mad-P signaling remains at high levels. This result suggests, together with results in the main text, that the Dpp signaling asymmetry seen in the haltere does not result from graded Dpp production, but from an anterior bias in Dpp diffusion. (D) tkv-lacZ is repressed medially in the wing, but is expressed uniformly throughout the haltere (Crickmore and Mann, 2006; de Navas et al., 2006; Makhijani et al., 2006). (E) DV wg expression is repressed in the P compartment of the haltere (arrow) (Weatherbee et al., 1998). (F) ptc-Gal4 expression is relatively uniform throughout its expression domain in the wing and haltere. (G) Intensity trace showing ptc-Gal4; UAS-GFP reporter expression in the boxed region of the haltere. ptc-Gal4 activity is only slightly graded at the anterior boundary of expression, arguing that the asymmetric extracellular Dpp::GFP detected in halteres of ptc-Gal4; UAS-dpp::GFP flies is due to asymmetric diffusion, not production. (H) Dlp protein levels are generally lower throughout the haltere when compared with the wing. Also, the haltere lacks the domain of DV Dlp reduction seen in the wing. (I) Hh protein is detected at much lower levels in the P compartment of the haltere (arrow) when compared to the wing.

• Supplemental Figure 2 -

  Fig. S2. The extracellular Dpp::GFP gradient in the wing. (A-A′′′) A wing disc expressing UAS-dpp::GFP with ptc-Gal4. Dpp diffuses well through the wing to form broad ligand and activity gradients on both sides of the AP organizer. This diffusion is slightly hampered in the P compartment due to a small increase in tkv levels in this compartment (Tanimoto et al., 2000). (B) Intensity traces from the disc shown in A.

• Supplemental Figure 3 -

  Fig. S3. The dally expression pattern. (A) Expression of the dally-lacZ enhancer trap in the wing. (B) An in situ hybridization for dally transcripts in the wing reveals a very similar expression pattern as is seen with dally-lacZ (see also Fujise et al., 2001). (C) Expression of dally-lacZ enhancer trap in the haltere. Arrows mark points of reference for comparison with D. (D) The pattern of dally mRNA in the haltere is similar to that reported by dally-lacZ. Note the decreased dally mRNA levels in the P compartment (red arrow) and increased staining laterally in the A compartment (yellow arrow).

• Supplemental Figure 4 -

  Fig. S4. Alterations in morphogen signaling resulting from changes in dally expression. (A,A′) Mad-P is reduced in dally mutant clones (arrow, marked by the absence of GFP) (see also Belenkaya et al., 2004). (B,B′) Mad-P is increased in dally⁺ clones (arrows, marked positively by GFP) (see also Fujise et al., 2003). (C,C′) Clones of cells in which Dally is knocked down using UAS-dallyRNAi (arrows, marked positively by GFP) show reduced Hh staining. (D,D′) Wg staining is increased in cells expressing high dally levels. Here, ci-Gal4 is used to drive UAS-dally^strong in A cells (marked by GFP) resulting in stronger Wg detection in the A compartment (see also Han et al., 2005). A white line indicates the AP boundary.

• Supplemental Figure 5 -

  Fig. S5. Intensity traces of dally-overexpressing halteres. These discs were stained together and imaged identically, and are therefore directly comparable. (A) An en>GFP haltere disc stained for Mad-P. The white arrow indicates the approximate position of the AP boundary. (B) An en>dally haltere disc stained for Mad-P. The white arrow indicates the approximate position of the AP boundary. (C) Intensity plots of A and B. A clear drop in Mad-P staining is seen in A cells adjacent to the AP boundary in the en>dally haltere disc.

• Supplemental Figure 6 -

  Fig. S6. Dpp target gene responses to dally overexpression. (A,A′) In the wild-type haltere, the Dpp target omb (reported here by omb-lacZ) is expressed mostly in the A compartment. (B,B′) Driving Dpp into the P compartment by expressing UAS-dally^strong with en-Gal4 (marked by GFP) increases the extent of omb-lacZ detection in posterior haltere cells.