Figure 6. JAK/STAT signaling is important for the formation of the ectopic wings.

(A) Frequency of ectopic wings (EW) following rn^ts> driven expression of >upd1, >egr or >upd1 together with >egr in wild type and CtBP^Q229*/+discs. Data were compared using ANOVA followed by Tukey test for significance (*p<0.05, **p<0.01, ***p<0.001) (B) Wing disc following rn^ts> driven co-expression of >egr and >upd1 after 72 hr of recovery. Pouch identity is shown by anti-Nub and an asterisk marks the ectopic pouch. (C–D) Early L3 (C) and late L3 (D) wing discs stained with anti-Wg and anti-GFP to detect the fast turnover STAT-DGFP reporter. (E–F) Wing discs with STAT-DGFP reporter following rn^ts>egr damage at 0 hr of recovery in wild type (E) and CtBP^Q229*/+ (F) stained with anti-MMP1. Note expression of MMP1 in a subset of cells expressing STAT-DGFP in the area of ectopic pouch formation (asterisk); see (Figure 6—figure supplement 1). (G–I) Wing imaginal discs with both upd3-lacZ and STAT-DGFP reporters in undamaged (G) and damaged wild type (H) and CtBP^Q229*/+ (I) discs at 0 hr of recovery. Note area in the notum that expresses STAT-DGFP but does not have elevated levels of upd3-lacZ. (J) Frequency of EW following rn^ts>egr damage when crossed to 1) FRT82B, 2) FRT82B Stat92E^85C9, 3) FRT82B CtBP^Q229*, and 4) FRT82B CtBP^Q229* Stat92E^85C9. Data were compared using ANOVA followed by Tukey test for significance (*p<0.05, **p<0.01, ***p<0.001). (K) Frequency of EW following rn^ts>egr damage in wild type and CtBP^334Δ4/+ in the listed genotypes (1) +/+, (2) upd1^YM55/+, (3) upd2^Δ/+, (4) upd3^Δ/+, and (5) upd2^Δupd3^Δ/+. In addition, co-expression of >upd1 with rn^ts>egr in (1) CtBP^334Δ4/+ and (2) upd2^Δupd3^Δ/+;; CtBP^334Δ4/+ genetic backgrounds. Note the rescue of EW induction in upd2^Δupd3^Δ/+ background by the co-expression of >upd1.

Figure 6—figure supplement 1. STAT-DGFP expression in the notum of damaged wing discs is independent of MMP1 expression.

(A–D) Multiple examples of discs with variable amounts of STAT-DGFP and MMP1 expression in the notum. The green arrowheads (A–A’’’) point to a region in the notum with STAT-DGFP expression without detectable MMP1 protein. The white arrowheads (B–C’’’) show an example of a large patch of STAT-DGFP expression with only small region of MMP1 expression. The red arrowheads (D–D’’’) show a high level of MMP1 expression within a region of STAT-DGFP expression. Note that the STAT-DGFP expression appears lower where it overlaps with MMP1 expression.

Figure 6—figure supplement 2. Upd1 overexpressed in the myoblasts can act on the disc proper epithelium and disrupt the notum Wg stripe.

Wing discs with STAT-GFP reporter in wild type (A–B) and CtBP^Q229*/+ (C–D) genetic backgrounds. (A, C) Control discs. (B, D) Discs with myoblast driver, R15B03-GAL4, expressing UAS-upd1. Note the disruption of the normal notum Wg stripe (arrows) and the expression of STAT-GFP reporter in the notum epithelium. (WP >RFP = R15C03-lexA, lexAOp-RFP is used to visualize the wing pouch). (E) Adult R15B03>upd1 flies. Note the disruption of the thorax including the loss of many bristles and macrochaetes (arrow).

Figure 6—figure supplement 3. CtBP^-/- mutant clones do not alter expression of upd3-lacZ.

(A) The upd3-lacZ reporter in CtBP^-/- mutant clones, which is marked by the absence of RFP (shown in magenta), in a wing disc. Note that upd3-lacZ is not expressed in the late L3 wing disc during normal development and that no expression was detected in CtBP^-/- mutant clones.

Figure 6—figure supplement 4. Effect of reducing CtBP function on STAT activity.

Wing discs with homozygous mutant clones of CtBP^Q229* with the STAT-DGFP reporter. CtBP^-/- mutant clones are marked by the absence of RFP. Areas in the white dotted box in (A) and (C) are enlarged in the subsequent panels (B–B”) and (D–D”).