(a) Representative confocal images of the prothoracic neuromere (PN) region two days after the front right leg injury are shown. GFP-labeled Gr22c gustatory axons in the PN region of control flies (left), with co-expression of Wlds (center), or in a draper mutant background (right). Arrows point to axon that failed to degenerate in Wlds-expressing axons or degenerating axons that remained uncleared in draper mutants. (b) Representative one micron images of adult VNCs from flies that carry the AP-1 reporter TRE-eGFP, 10XSTAT92E-dGFP, or the Draper reporter dee7-GFP. Reporter activation was largely restricted to the PN and AMN regions after injury to both front legs and wings. Dotted line shows outline of the posterior VNC tissue housing uninjured projections. (c) Quantitative real-time PCR of normalized expression levels of draper-I transcript in VNCs following injury to all legs and both wings (magenta asterisks) or following injury to all legs, both wings, and head decapitation (blue circles). Draper threshold cycle (Ct) values were normalized to ribosomal protein L32. Biological replicates: 6 legs + 2 wings: No Injury N = 8; 1.5 hr N = 3; 5 hr N = 3. 6 legs + 2 wings+head: No Injury N = 8; 5 hr N = 7. Mean ± SEM plotted; ****p<0.0001; Two-way ANOVA with Sidak post hoc test. Each injury group was compared to uninjured in the same group. Black asterisks depict comparison between the two injury groups at the 5 hr time point. Scale bars = 30 μm. Genotypes: Figure 3a: Control: Gr22c-Gal4/+; UAS-mCD8::GFP/+. Wlds: UAS-Wlds/+; Gr22c-Gal4/+; UAS-mCD8::GFP/+. draper-/-: Gr22c-Gal4/UAS-mCD8::GFP; draperΔ5rec9/draperΔ5rec9. Figure 3b: TRE-eGFP: TRE-eGFP/TRE-eGFP (on II); 10xSTAT92E-dGFP: 10xSTAT92E-dGFP/10xSTAT92E-dGFP (on II); dee7-Gal4, mGFP: dee7-Gal4, UAS-mCD8::GFP/CyO. Figure 3c: w1118.