DEV014670 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Brk, Wg, P-Mad and Dll expression domains in early stages of leg development. (A) A second instar leg disc (∼60 hours AEL) stained for Dll (green), P-Mad (blue) and Brk-lacZ (red). Dll is present in the center of the disc where there is P-Mad, but no detectable Brk-lacZ. (B) At the beginning of the third instar (∼72 hours AEL), the Dpp (visualized by P-Mad staining, blue) and Wg (green) gradients are restricted to dorsal and ventral sectors, respectively. brk expression is restricted to the ventral and lateral disc.

• Supplemental Figure S2 -

  Fig. S2. brk is negatively regulated by Dpp signaling. (A) Positively marked clones expressing a constitutively activated form of the Dpp receptor Tkv^QD (green, arrow) cell-autonomously repressed brk (red). (B) Elimination of Dpp activity in Mad loss-of-function clones (negatively marked by the absence of GFP, green) derepressed brk-lacZ (red; arrow).

• Supplemental Figure S3 -

  Fig. S3. brk activity is not required for DV axis specification. (A,B) Wild-type pattern of Dpp pathway activation (monitored by P-Mad staining; (A) and wild-type pattern of Wg (B) in 3rd instar leg discs. (C,D) In brk^XA discs, the P-Mad (C) and Wg (D) patterns are dorsally and ventrally restricted, respectively, as in wild type. These 3rd instar discs are overgrown relative to wild type. (E,F) In brk^XA; dpp^discs discs, there is no detectable P-Mad staining (E) and, consistently, Wg is derepressed dorsally (F).

• Supplemental Figure S4 -

  Fig. S4. Dpp is required for DV specification. (A-C) Positively marked clones expressing a constitutively activated form of the Dpp receptor Tkv^QD (green, arrow) cell-autonomously repressed ventral molecular markers (red) H15 (A,A′) and wg (B,B′). (D,E) yellow-marked clones expressing a constitutively activated form of the Dpp receptor Tkv^QD generate dorsal identities in the ventral adult leg. (D) A ventral Tkv^QD clone in the femur transforms ventral identities towards dorsal, as observed by the appearance of dorsal bristles (arrow) in the ventral femur. (E) A ventral Tkv^QD clone in the first tarsus creates PD duplications and promotes dorsal identities (arrow). (F) brk⁻ mutant clones in the distal leg (distal tibia and tarsi) have no phenotype (arrow).