Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Rabbit Poly(mono)clonal α-Oskar | A. Ephrussi | N/A | 1:1000 or 1:2500 (see Materials and methods) |
| Antibody | Rabbit polyclonal α-Vasa | R. Lehmann | N/A | 1:1000 |
| Antibody | Mouse monoclonal α-Lamin | Developmental Studies Hybridoma Bank | ADL84.12 | 1:1000 |
| Antibody | Alexa Fluor 488 Phalloidin | Molecular Probes | A12379 | 1:500 |
| Antibody | Rabbit polyclonal α-mCherry | Abcam | Ab167453 | 1:1000 |
| Antibody | Goat α-rabbit HRP | Abcam | Ab6721 | 1:2000 |
| Antibody | Goat α-rabbit horse HRP | Jackson ImmunoResearch | 111-035-003 | 1:5000 |
| Antibody | Mouse α -mouse HRP | Rockland | 18-8817-31 | 1:5000 |
| Antibody | Mouse monoclonal α-alpha tubulin | Sigma-Aldrich | T9026-2ML | 1:10000 |
| Antibody | Rabbit α-Aubergine | R. Lehmann | N/A | 1:1000 |
| Antibody | Rabbit α-Ser2 | Abcam | Ab5095 | 1:750 |
| Chemical | Vectashield Antifade Mounting medium | Vector Labs | Cat# H-1200 | |
| Chemical | Prolong Diamond Mounting medium | Molecular Probes | Cat# P36961 | |
| Chemical | Deionized formamide | Life Technologies | Cat# 4311320 | |
| Chemical | 10% Fetal Bovine Serum | Invitrogen | Cat#10082-139 | |
| Chemical | Penicillin-streptomycin | Fisher | Cat# 15140122 | |
| Chemical | Poly-L-lysine solution | Sigma | Cat# P4707-50ml | |
| Chemical | 20% Paraformaldehyde | Electron Microscopy Sciences | Cat#15713 | |
| Chemical | BSA | Sigma | Cat#A4503-100G | |
| Chemical | Restore Western Blot Stripping Buffer | Fisher Scientific | Cat# PI21059 | |
| Chemical | Triton X-100 | Sigma-Aldrich | Cat# T8787-250ML | |
| Chemical | 1,6 Hexanediol | Sigma-Aldrich | Cat#240117-50G | |
| Commercial assay | NuPAGE Novex 4-12% Bis-Tris Gel 1.0mm, 10 well | Invitrogen | Cat# NP0321BOX | |
| Commercial assay | Immun-Blot PVDF Membrane | Bio-Rad | Cat# 162-0174 | |
| Commercial assay | Imaging membranes | YSI | Standard | |
| Commercial assay | Effectine Transfection reagent | Qiagen | Cat# 301427 | |
| Commercial assay | Gibson Assembly Master Mix | New England BioLabs | Cat#E2611L | |
| Commercial assay | Nunc Lab-Tek II Chambered Coverglass | Thermo Fisher Scientific | Cat# 155409PK | |
| Cell line | D. melanogaster S2R+ | Drosophila Genomics Resource Center | FlyBase: FBtc0000150 | |
| Cell line | Human:HEK293T | ATCC | 293T (ATCCCRL3216TM) | |
| Organisms/Strains | Wild-type:w-1118 | Bloomington (BDSC) | Stock #3605 | |
| Organisms/Strains | y,w; P[E GFP-vas w+]cyIII | (Trcek et al., 2015) | N/A | |
| Organisms/Strains | UAS–vasa–ko | (Cinalli and Lehmann, 2013; Trcek et al., 2015 ) | N/A | |
| Organisms/Strains | pFlyFos-Osk/CyO | (Jambor et al., 2015; Trcek et al., 2015) | N/A | |
| Organisms/Strains | UASp-GFP-Aub | (Webster et al., 2015) | N/A | |
| Organisms/Strains | GFP-Piwi | (Le Thomas et al., 2013) | N/A | |
| Organisms/Strains | GFP-Tud | R. Lehmann | N/A | |
| Organisms/Strains | ;;P{GAL4::VP16-nos.UTR}/Tm3Ser | R. Lehmann | N/A | |
| Organisms/Strains | w[*]; P{w[+mC]=His2Av-mRFP1}II.2 | Bloomington (BDSC) | Stock #23651 | |
| Organisms/Strains | w;;OskΔNLS-alleleA/TM3 | This study | N/A | See Materials and methods |
| Organisms/Strains | w;;OskΔNLS-alleleB/TM3 | This study | N/A | See Materials and methods |
| Organisms/Strains | w;His2AAv-mRFP/cyo;OskΔNLS-alleleA,Vasa:GFP/TM3 | This study | N/A | See Materials and methods |
| Organisms/Strains | w; matα-gal4;PrDr/TM3 | (Pae et al., 2017) | N/A | |
| Organisms/Strains | Df(3R)Pxt103/TM3 Ser Sb | (Lehmann and Nüsslein-Volhard, 1991) | N/A | |
| Organisms/Strains | osk54 st ry ss/TM3Ser | (Lehmann and Nüsslein-Volhard, 1991) | N/A | |
| Organisms/Strains | w; P(EPgy2)gclEY09611/Cyo; nosGal4VP16 (w+)/TM3 | (Cinalli and Lehmann, 2013) | N/A | |
| Organisms/Strains | FT553: zuIs247 [nmy-2::PGL-1-GFP line O05-2]; unc-119(ed3) III; ltIs44 V [pie-1p-mCherry::PH(PLC1delta1)] | J. Nance | N/A | |
| Oligonucleotides | Osk nls gRNA-A | This study | N/A | See Materials and methods |
| Oligonucleotides | Osk nls gRNA-B | This study | N/A | See Materials and methods |
| Oligonucleotides | ssODN for gcl-NLS-A | This study | N/A | See Materials and methods |
| Oligonucleotides | ssODN for gcl-NLS-B | This study | N/A | See Materials and methods |
| Oligonucleotides | Calca Fluor 590-conjugated nanos smFISH probes | (Trcek et al., 2015) | N/A | |
| Oligonucleotides | Quasar 670-conjugated gcl smFISH probes | (Trcek et al., 2017) | N/A | |
| Oligonucleotides | Quasar 670-conjugated cycB smFISH probes (5′→3′): cgttttgttgttgcctccat, tatgccgcgattctgcaaat,tctttctgtgccgcatcctt, tctgtgagcttgagatcctt, tccacccgagctttggcatt, agtggctgtttcttccagtg, ttgccattgccattggtgct,aattcgaacgcaaaaacgcc, tacagtggtcttggtcggaa,tgactgtaactttagtgggt, ttcacgttctcggaagaact,ttgctgtcctcgcgctttaa, agtttggtcagcgacttctt,attcctgaaactcccatcac, aaacagctactggttcccgt,ttcttggtctctgcctcttt, cttcttggtttctggcagtt,cctttttcacttccagtgag, tgcactgttgcccctaataa, atcgtagatgtggtcgtagt,ttgctggaaagggacatggt, atcaatgtcctcgattccag,accaggttctccttgtcatt, cgtttacatattcggagacc, cctgatacaagtagtcgtag, atccttgtgaatgggctgct,atcttgtgggacacctcctt, ttgatccaatcgatcagcac,atggaactgcaggtggactt, tagcgatcaatgatagccac, tttggtgtccttgaccacct, actcccaccaattgcaagta,acttggtggctatgaagagt, aagacgaaatctccgattgc,agtgtaggtgtcgtccgtga, agatttgcagctccatctgt,agattacagtcgatggcctt, tagcgtcgaaggaagtgaat,ttggacatcgtatggtgctc, ggaagctaactcgatgaagt,taagtggccatttcgtagtc, aacagtgaggcagctgcaat,tttccattgagcaagtgcag, acggtcgttgaatcctgtac,atcgcgagtagaaggtcaga, ttcgcgatcagccgggtaat,ttgtagatggccttcagctt, atcttctggaacttgctgcc |
This study | N/A | See Materials and methods |
| Recombinant DNA | pActin5C | Drosophila Genomics Resource Center | Cat#1112 | |
| Recombinant DNA | pVERMILION22 | (Ni et al., 2011) | N/A | |
| Recombinant DNA | pEF1/V5-His A | ThermoFisher Scientific | V92020 | |
| Recombinant DNA | pDendra2 | Addgene | Cat#75283 | |
| Software and Algorithms | ImageJ | NIH | https://imagej.nih.gov/ij/ | |
| Software and Algorithms | NLS finder | cNLS Mapper | http://nls-mapper.iab.keio.ac.jp/cgi-bin/NLS_Mapper_form.cgi | |
| Software and Algorithms | IDR and LC prediction | SEG IUPRED JPRED |
http://mendel.imp.ac.at/METHODS/seg.server.html
http://iupred.enzim.hu/ http://www.compbio.dundee.ac.uk/jpred/ |
|
| Software and Algorithms | EasyFRAP | (Rapsomaniki et al., 2012) | N/A | |
| Software and Algorithms | Huygens deconvolution software | Scientific Volume Imaging | N/A | |
| Software and Algorithms | Prism 7 | Graphpad | N/A | |
| Software and Algorithms | SigmaPlot | SigmaPlot (Systat Software, Inc) | N/A | |
| Software and Algorithms | CRISPR design for guide RNAs | MIT | http://crispr.mit.edu/ | |
| Software and Algorithms | CRISPR design for guide RNAs | ZiFit | http://zifit.partners.org/ZiFiT/ | |
| Software and Algorithms | DAVID GO term analysis | The Database for Annotation, Visualization andIntegrated Discovery (DAVID) version 6.7. | https://david.ncifcrf.gov/ |