Uas egfp

Flies were kept at 25°C. The following stocks/mutant alleles were used: OregonR as wild-type control, crb^11A22[30] , crb^GX24[31] (link), crb^8F105[30] , [32] (link)
foscrb_Y10A,ΔERLI; crb^GX24[33] (link), UAS-crb^30.12e[14] (link) called UAS-crb^full here, en-Gal4 [34] (link). Mutant stocks were balanced over TM3, Twist-Gal4, UAS-EGFP (Bloomington Stock Center).

Fly stocks were raised at 18 °C, and all the experiments were performed at 25 °C (12 h daytime and 12 h nighttime, 60% humidity) on standard cornmeal food. If necessary, the food was supplemented with other chemicals. Fly stocks Actin-GAL4/CyO (Bloomington #4414), Da-GAL4 (Bloomington #8641), and UAS-EGFP (Bloomington#5130) were obtained from the Bloomington Stock Center (Bloomington, IN, USA); NP3084-GAL4 (DGRC# 113094) was from the Drosophila Genetic Resource Center at the Kyoto Institute of Technology (Kyoto, Japan). Esg-GAL4, pros-GAL4, and myo1A-GAL4 are a gift from Wei Zhang lab (Tsinghua University, China). The RNAi lines used for the screen were attached in the supplement Additional file 1: Table S1.
Hemin was dissolved in DMSO at 100 mM. 4,6-Dioxoheptanoic acid (succinylacetone, SA) was dissolved in H₂O at 50 mg/ml, and the zinc-mesoporphyrin (Logan, UT) was prepared in DMSO at 100 mM.

All Drosophila stocks were maintained on standard cornmeal at 25 °C in light/dark-controlled incubators. The w1118, UAS-eGFP, and D42-GAL4 were obtained from the Bloomington stock center. The UAS-FUS WT, UAS-FUS P525L, and UAS-FUS R521C flies as well as the experimental conditions for climbing index assessment were previously described [1 (link)].

w¹¹¹⁸, P{EP}Nup44A^EP2417, P{EPg}Nup93-2^HP35056, P{UAS-NLS-NES-GFP}, P{UAS-NLS-ΔNES-GFP}, Ok371-gal4, and UAS-eGFP were obtained from the Bloomington Drosophila stock center (http://www.flystocks.bio.indiana.edu). UAS-Nup214 (F001467) and UAS-Nup43 (F003133) were obtained from FlyORF. Nup62 RNAi (100588) and Nup214 RNAi (41964) were obtained from Vienna Drosophila Resource Center. The previously described CRISPR/Cas9 human TDP-43 wild-type (hTDP-43WT) was a kind gift from David B. Morton (Oregon Health and Science University) (Chang and Morton, 2017 (link)). UAS-Nup62 overexpression lines (UAS-Nup62 OE) were generated through site-specific integration of the transgene at BestGene Inc To generate UAS-Nup62 OE lines, we used the pUAST-attB vector. We subcloned the Nup62 cDNA (Drosophila Genomic Research Center) into the pUAST-attB vector.

The following mutant and transgenic flies were used in this study: GMR-Gal4 (Bloomington Stock Center), UAS-EGFP (Bloomington Stock Center), UAS-AR14Q (current study), UAS-AR61Q (current study), UAS-trAR^112Q (Chan et al., 2002 (link)), UAS-NLK-WT (Ju et al., 2013 (link)), UAS-NLK-KN (Ju et al., 2013 (link)), nmo^adk1 (Verheyen et al., 2001 (link)), nmo^adk2 (Verheyen et al., 2001 (link)). In order to generate UAS-AR14Q and UAS-AR61Q transgenic fly lines, full-length human AR cDNAs with 14Q or 61Q were subcloned into the pUAST vector and then injected into fly embryos (via Best Gene, Inc. Chino Hills, CA). After crossing with the GMR-Gal4 driver line, two independent UAS-AR lines of each Q length that showed roughly equal levels of transgene expression were used for the analysis. For the genetic interaction analyses, appropriate fly lines were intercrossed and their progeny were raised at 22°C, 25°C, or 30°C on fly food containing or lacking 100 nM DHT. All experiments were carried out multiple times.

Fly stocks were raised at 18 °C, and all the experiments were performed at 25 °C on standard cornmeal food. When necessary, the food was supplemented with metals or metal chelators at stated concentrations. Fly stocks act-GAL4/CyO (Bloomington #4414), da-GAL4 (Bloomington #8641) and UAS-EGFP (Bloomington#5130) were obtained from the Bloomington Stock Center (Bloomington, IN, USA); NP3084 (DGRC# 113094) and NP1093 (DGRC# 103880) lines were obtained from the Drosophila Genetic Resource Center at the Kyoto Institute of Technology (Kyoto, Japan). The MtnB-EYFP line was a kind gift from Dr. Walter Schaffner (University of Zurich, Zurich, Switzerland). The RNAi lines were provided by the VDRC (Vienna, Austria) or custom made at the Tsinghua Fly Center (Beijing, China), Zip71B-RNAi (VDRC#44539 and TH00155.N), ZnT35C-RNAi (VDRC#103263 and TH00119.N), ZnT41F-RNAi (VDRC#5390), and Xdh-RNAi (THU5626). Transgenic flies were generated in w¹¹¹⁸ background by P-element-mediated transformation.