Drosophila Genetic Toolbox for Protein Localization

The P(Mae-UAS.6.11)^{23 (link)}, P(BacWH)^{24 (link)}, P(EP)²⁵²², and P(EPg)^{25 (link)} enhancer-promoter lines on the X-chromosome were obtained from the Bloomington Drosophila stock center (BDSC). The following EP lines, inserted in the direction of the Fim gene expression were screened for eye phenotypes: P(XP)Fim^d02114, P(XP)Fim^d05016, P(XP)Fim^d03334, P(EP)Fim^G10929. In addition, we used the following BDSC lines: the Fim protein trap line P(PTT-GC)Fim^CC01493, UAS-Syt::eGFP (BDSC_6926), UAS-Lifeact::Ruby (BDSC_35545), UAS-Act5C::GFP (BDSC_9258), UAS-mCD8::RFP (BDSC_32218), UAS-Pod1 (BDSC_8800), P(EPgy2)coro^EY05114 (BDSC_19703), deficiency line covering Fim (Fim^Def) is BDSC_4741, and the Fim^XP (BDSC_ 19171) and Fim^e03892 stock was obtained from the Exelixis collection. UAS-RNAi lines, including the one used for IKKε (VDRC_103748) were from the Vienna Drosophila Research Center. The following Gal4 lines were used: GMR-Gal4, nSyb-Gal4, A307-Gal4. For the FRAP assay, nSyb-Gal4 was recombined with UAS-Syt::eGFP on the third chromosome. The insertion sites of the EPs were determined by inverse PCR following the protocol indicated in the Gene Disruption Project (http://flypush.imgen.bcm.tmc.edu/pscreen/). Unless otherwise stated, we assessed female flies throughout the experiments.