Increased expression of tribbles (trbl) causes motor impairment and decreases survival. (A) Analysis of Trbl expression in larval eye discs. Third instar eye imaginal discs of UAS-trbl-HA flies driven by gmr-Gal4, immunostained for HA (green) and Hoechst (purple). (B) Ubiquitous expression of the UAS-trbl-HA leads to increased trbl transcript levels. mRNA levels were measured by real-time qPCR (mean+s.e.m.; ***P<0.001; unpaired t-test; n=4 biological replicates). (C) Ubiquitous expression of trbl causes a delay in development, with reduced numbers of developed pupae (mean+s.e.m.; **P<0.01; unpaired t-test). (D,E) Ubiquitous expression of trbl leads to transcriptional upregulation of Thor (D) (mean+s.e.m.; ****P<0.0001; unpaired t-test; n=4 biological replicates), but not InR (E) (mean+s.e.m.; NS, not significant; unpaired t-test; n=4 biological replicates). Transcripts were measured in 14-day-old adult males. (F) Ubiquitous expression of trbl decreases motor performance. Adult flies were tested using a standard climbing assay (mean±s.e.m.; *P<0.05; two-way ANOVA with Sidak multiple comparisons test; n≥5 biological replicates). (G) Ubiquitous trbl expression reduces lifespan (log-rank test). Genotypes: in A, w; gmrGal4/UAStrbl-HA;+; in B-G, w; +; daGal4/+ (control), w; UAStrbl-HA/+; daGal4/+ (UAS-trbl).