Figure 7. ube3a was required to form camphor-induced taste desensitization.

(a) Behavioral responses to 1 mM sucrose versus 5 mM sucrose plus 6 mM camphor. The tests were performed using wild-type flies, ube3a^15b, or the mutant animals harboring a genomic ube3a⁺ rescue transgene. The animals were fed a normal diet, a camphor diet for 24 hours, or a camphor diet followed by returning to a normal diet for 24 hours. n=5 trials. *p=0.00082 (WT). (b and c) TRPL protein levels in the proboscis of wild-type, ube3a^15b and rescued flies after feeding a normal or camphor diet. The full-length blots are shown in Supplementary Fig 10d. n=3 trials. *p=0.00078 (WT). (d and e) Tip recordings showing responses of S6 sensilla to 6 mM camphor. The recordings were performed using wild type, ube3a^15b and ube3a^15b flies containing the genomic ube3a⁺ rescue transgene (rescue). The animals were maintained on a normal diet, a camphor diet for 24 hours, or a camphor diet for 24 hours and then a camphor-free diet for 24 hours. n=8 animals. *p=0.0022 (WT). (f-h) Immunocytochemical analysis of TRPL expression in a TRPL::GFP S2 cell line transfected with a vector encoding Ube3a with an N-terminal Myc tag (Myc::Ube3a). Myc::Ube3a and TRPL::GFP expression was revealed by anti-Myc and anti-GFP staining, respectively. The scale bar indicates 10 μm. (i) Quantification of the fluorescence intensity of TRPL::GFP in the TRPL::GFP S2 cell line transiently-expressing Myc::Ube3a. n=3 trials with ~10 cells for each trial. **p=0.00013. (j) Cell lysates of the TRPL::GFP S2 cell line with or without Ube3a::Myc. TRPL::GPF was immunoprecipitated with anti-TRPL and the blot was probed with anti-Ubiquitin. The loading controls were blotted with anti-Myc and anti-TRPL. Error bars indicate SEMs. One-way ANOVA tests with Bonferroni post-hoc analysis.