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En gal4

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En-Gal4 is a lab equipment product that serves as a tool for gene expression analysis. It functions as a genetic construct that allows for the targeted expression of genes in specific cell types or developmental stages. The En-Gal4 product provides a means to study gene function and regulation in a variety of experimental systems.

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Lab products found in correlation

Ptc gal4, by BD (3 mentions) Gmr gal4, by BD (2 mentions) Ey gal4, by BD (2 mentions) Gmr gal4, by Bloomington Drosophila Stock Center (1 mentions) Uas p35, by BD (1 mentions) Uas diap1, by BD (1 mentions) Uas cyce, by BD (1 mentions) Nub gal4, by BD (1 mentions) En gal4, by Bloomington Drosophila Stock Center (1 mentions) Uas rfp, by BD (1 mentions) Nre gfp, by Bloomington Drosophila Stock Center (1 mentions) Uas br rnai, by BD (1 mentions) Nre gfp, by BD (1 mentions) Uas bskdn, by BD (1 mentions) Apgal4, by BD (1 mentions) Dpp gal4, by BD (1 mentions) C96 gal4, by BD (1 mentions) Act5c gal4, by BD (1 mentions) Elav gal4, by BD (1 mentions) Mef2 gal4, by BD (1 mentions)

8 protocols using en gal4

1

Drosophila Genetics Protocol

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All Drosophila strains were grown and maintained at 25 °C. Following fly stocks were used for the experiments: Ciao1 RNAi (Vienna Drosophila Resource Center, v32020 and v105939), Xpd RNAi (Vienna Drosophila Resource Center v106998), UAS-CycE (BDSC 4781), UAS-Diap1, UAS-p35, ey-Gal4, GMR-Gal4, en-Gal4, ptc-Gal4, nub-Gal4, and FRT42D M(2)531 (BDSC 5698) were obtained from the Bloomington stock center. For overexpression of Crbintra, UAS-Crbintra was crossed with GMR-Gal4 (Bloomington). Xpdp flies were a kind donation from Dr. Beat Suter. To construct the transgenic lines, UAS-Ciao1 and UAS-Xpd, full length Ciao1 and Xpd cDNA (from the Drosophila Genome Research Center) were cloned into pUAST vector.
Jung J., Yeom E, & Choi K.W. (2020). Ciao1 interacts with Crumbs and Xpd to regulate organ growth in Drosophila. Cell Death & Disease, 11(5), 365.
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2

Drosophila Genetic Manipulation Protocol

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Strains contain FRT 19A brnpr-3 and hsFLP, FRT 19A RFP (Jia et al., 2014 (link)). The following transgenic lines were used in our study: en-Gal4, UAS-mRFP, NRE-GFP (Bloomington Drosophila Stock Center, BDSC#30729, USA), en-Gal4, UAS-mRFP, NRE-GFP; Gal80ts, UAS-br RNAi (BDSC#27272), UAS-mam RNAi (BDSC#28046), UAS-EcR B1 W650A (BDSC#6872), UAS-br-Z1 (Zhou and Riddiford, 2002 (link)), UAS-Delta RNAi (BDSC#34322), UAS-Delta (BDSC#5614), UAS-NICD (Xie et al., 2014 (link)), cut-lacZ (Jack et al., 1991 (link)) and hsFLP; actin<CD2<Gal4,UAS-RFP/TM3,Sb.
For the FLP/FRT clone induction (Golic and Lindquist, 1989 (link); Xu and Rubin, 1993 (link)), hsFLP, 19A RFP was applied, and clones were induced by one session of heat shock for 2 h at 37°C during late second instar larval stage. To generate mosaic wing discs expressing UAS constructs, the flip-out Gal4 (Pignoni and Zipursky, 1997 (link)) stock hsFLP;actin<CD2<Gal4,UAS-RFP/TM3,Sb was applied. Flip-out clones were induced by 30 min heat shock at 37°C two days before dissection. en-Gal4,UAS-mRFP, NRE-GFP; Gal80ts was used to manipulate gene expression in the posterior compartment of wing discs. In this technique, en-Gal4,UAS-mRFP, NRE-GFP; Gal80ts flies were crossed with UAS lines, and the progeny were raised at 18°C until late second instar larval stage, then shifted to 29°C for two days before dissection.
Jia D., Bryant J., Jevitt A., Calvin G, & Deng W.M. (2016). The Ecdysone and Notch Pathways Synergistically Regulate Cut at the Dorsal-Ventral Boundary in Drosophila Wing Discs. Journal of genetics and genomics = Yi chuan xue bao, 43(4), 179-186.
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3

Drosophila Genetics and Imaging Protocols

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All fly stocks were maintained on standard cornmeal/yeast/molasses/agar medium at 25 °C as per standard procedures. Oregon-R flies were used as wild-type controls. UAS-GFP-zip, UAS-GFP-Zip DN (UAS-Myo II-Neck-Rod), UAS-Myo II-Rod, and UAS-Myo II-Rod (delta Nterm58) [26 (link)] were obtained as a gift from Prof. Daniel P. Kiehart (Department of Biology, Duke University, Durham, NC). UAS-Notch-FL [27 (link)], UAS-Notch-ICD, UAS-Notch-DN [28 (link)], and Notch pathway components were kindly provided by Prof. S. Artavanis-Tsakonas (Department of Cell Biology, Harvard Medical School, Boston, MA).

cn1 bw1 sp1 zip1/CyO (BDSC 4199),

P{FRT(whs)}G13 zip2/CyO (BDSC 8739),

P{lacZ.w+}276, y1 sc* v1 sev21; P{TRiP.GL00623}attP40 (BDSC 37480), y1 sc* v1 sev21; P{TRiP.HMS01618}attP2 (BDSC 65947),

SqhAX3-GFP (BDSC 57144), UAS-Bsk-DN (BDSC 6409), vg-GAL4 (BDSC 8222), GMR-GAL4 (BDSC 8121), ptc-GAL4 (BDSC 2017), en-GAL4 (BDSC 30564), ap-GAL4 (BDSC 56807), dpp-GAL4 (BDSC 1553), and C96-GAL4 (BDSC 43343) stocks were obtained from Bloomington stock center. All crosses were performed at 25 °C. The combination lines vg-GAL4/UAS-GFP-zip and UAS-Notch-DN/C96-GAL4 were made with the help of appropriate genetic crosses.

Verma D., Singh A., Singh J., Mutsuddi M, & Mukherjee A. (2024). Regulation of Notch signaling by non-muscle myosin II Zipper in Drosophila. Cellular and Molecular Life Sciences: CMLS, 81(1), 195.
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4

Fly Strains for Molecular Studies

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The following fly strains were used throughout the study: Act5C-Gal4 (BDSC #3954, Christian Klämbt), ey-Gal4 (BDSC #5535), ey-Gal4, UAS-Dl/CyO flies (a gift from B. Hassan), robo2-Gal4 (BDSC #48074), en-Gal4 (BDSC #6356), ptc-Gal4 (BDSC #52212), C179-Gal4 (BDSC #6450), mef2-Gal4 (BDSC #27390), CG25C-Gal4 (BDSC #7011), elav-Gal4 (BDSC #458), slit-Gal4 (BDSC #9580, Christian Klämbt), repo-Gal4 (BDSC #7415, Christian Klämbt), gcm-Gal4 (BDSC #35541), crol-Gal4 (VDRC #200123), ama-Gal4 (#205487), appl-Gal4 (a gift from Doris Kretzschmar, Münster) and tkk-Gal4 (#45606). All RNAi strains used are listed in (Table S6).
Chaouch A., Berlandi J., Chen C.C., Frey F., Badini S., Harutyunyan A.S., Chen X., Krug B., Hébert S., Jeibmann A., Lu C., Kleinman C.L., Hasselblatt M., Lasko P., Shirinian M, & Jabado N. (2021). Histone H3.3 K27M and K36M mutations de-repress transposable elements through perturbation of antagonistic chromatin marks. Molecular cell, 81(23), 4876-4890.e7.
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5

Drosophila Fly Stocks for Cell Biology

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The following fly stocks were used in this study: CFD2 and TH_attP2,70 (link) Snky-GFP,71 (link) GFP-LAMP,72 (link) GFP::TMF (this study, see below) αManII::GFP (this study, see below), GMAP_2.2.5, TMF_Δ19, golgin-84_ Δ13.3 (all three this study, see below), fkh-GAL4 (BDSC 78060), en-GAL4 (BDSC 1973), and YFP::pgant9 (75 (link), Kyoto Stock Centre 115535). The following UAS-X-mito stocks were generated (this study, see ΦC31 transgenesis): UASt-BioID2-mito (II), UASt-BioID2-mito (III), UASt-g245-mito (II), UASt-g245-mito (III), UASt-TMF-mito (II), UASt-TMF-mito (III), UASt-g84-mito (II), UASt-g84-mito (III), UASt-GMAP-mito (II), UASt-GMAP-mito (III).
Park S.Y., Muschalik N., Chadwick J, & Munro S. (2022). In vivo characterization of Drosophila golgins reveals redundancy and plasticity of vesicle capture at the Golgi apparatus. Current Biology, 32(21), 4549-4564.e6.
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6

Overexpression studies of Notch signaling components

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Overexpression studies were performed using the Gal4/UAS system36 (link). We used the following UAS lines: UAS-Dl, UAS-Ser, UAS-mCD8-ChRFP (Bloomington Drosophila Stock Center; BDSC), UAS-fwdIR, UAS-sktlIR, UAS-plc-21IR, UAS-dgkIR, UAS-gfpIR (Vienna Drosophila RNAi Center). UAS-pld38, and UAS-rdgA/dgk were a generous gift from Padinjat Raghu (National Centre for Biological Sciences, Tata Institute of Fundamental Research, India). We also used the following Gal4 lines: pnr-Gal4, en-Gal4 and sca-Gal4 (BDSC). We also used the following strains Dl[Rev10], Ser[RX82], SpdoG104, Ap-2α[40–31], numb1537 (link). SpdoCh2GFP3, neur-nlsFP670, NiGFP were a generous gift from François Schweisguth (Institut Pasteur, France). UAS-lipinwt was a generous gift from Michael Lehmann (University of Arkansas, USA). All phenotypes were analysed at 25 °C unless stated otherwise.
Medina-Yáñez I., Olivares G.H., Vega-Macaya F., Mlodzik M, & Olguín P. (2020). Phosphatidic acid increases Notch signalling by affecting Sanpodo trafficking during Drosophila sensory organ development. Scientific Reports, 10, 21731.
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7

Drosophila Lineage-Specific Gene Expression

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We used Oregon-R (Bloomington Drosophila Stock Center (BDSC) 6362) as a wild-type strain. For cell lineage specific gene expression, we utilized the Gal4-UAS system, in which transcriptional activator Gal4 is expressed in a specific cell lineage and binds to UAS (upstream activating sequence) to induce downstream gene expression. The following Gal4 lines were used: NP1-Gal4 (Kyoto stock center 112001), mex-Gal4 (a gift from Dr. Lucy Erin O'Brien), esg-Gal4 (a gift from Dr. Shigeo Hayashi), Dl-Gal4 (a gift from Dr. Xiankun Zeng), MS1096-Gal4 (BDSC 8860), and en-Gal4 (BDSC 25752). tub-Gal80 ts (BDSC 7019) was used to temporally regulate Gal4-driven gene expression. To induce mosaic gene expression, tub-FRT-Gal80 (BDSC 38880) and hs-FLP (BDSC 1929) were used. For the split-lacZ analysis, flies with the following genotype were used: hs-FLP; mex-Gal4, X15-33 and tub-Gal80ts, X15-29 Diptericin-lacZ reporter (BDSC 55707) was used to histologically examine the AMP expression by X-gal staining. Bloomington Deficiency Kit (BDSC, http://flystocks.bio.indiana.edu/ Browse/df/dfkit-info.htm) was utilized for the genome-wide screen to identify gene deletions that alleviate the mortality of NP1-Gal4-mediated Atg7 RNAi flies upon DSS feeding.
Nagai H., Tatara H., Tanaka-Furuhashi K., Kurata S, & Yano T. (2021). Homeostatic Regulation of ROS-Triggered Hippo-Yki Pathway via Autophagic Clearance of Ref(2)P/p62 in the Drosophila Intestine. Developmental cell, 56(1).
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8

Drosophila Maintenance and Genetic Stocks

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Flies were maintained at 25 C on medium containing malt 6.5% (w/v), semolina 3.2% (w/v), dry baker's yeast 1.8% (w/v), agar 0.6% (w/v), propionic acid 0.7% (v/v) and Nipagin (methylparaben) 2.4% (v/v) (Havula et al., 2013) or were grown on modified food containing 0.5% (w/v) agar, 2.5% (v/v) Nipagin (methylparaben) in PBS and supplemented with 20% (w/v) dry baker's yeast or the indicated concentrations of sucrose. Stocks used included UAS-Rheb (BDSC, 9688), UAS-InR (BDSC, 8262), UAS-Myc (Johnston et al., 1999) , UAS-Myc-RNAi (VDRC, 2947) TOR delP (BDSC, 7014), tub-Gal4 (Lee et al., 1999) , en-Gal4 (Fietz et al., 1995) , cg-Gal4 (BDSC, 7011), UAS-dPWP1-RNAi (NIG-Fly, 6751R-1 and 6751R-3), nclb 1 and nclb 2 (Casper et al., 2011) , HsFLP;;FRT82 GFP/ TM3, W;;Actin
Liu Y., Mattila J., Ventelä S., Yadav L., Zhang W., Lamichane N., Sundström J., Kauko O., Grénman R., Varjosalo M., Westermarck J, & Hietakangas V. (2017). PWP1 Mediates Nutrient-Dependent Growth Control through Nucleolar Regulation of Ribosomal Gene Expression. Developmental cell, 43(2).
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