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. 2018 Apr 23;45(2):212–225.e7. doi: 10.1016/j.devcel.2018.03.018
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Anti-LMN-1 Gift from Georg Krohne, University of Würzburg, Germany N/A
Anti-LMN-1Ser32pi This paper N/A
Anti-SUN-1Ser8pi (Penkner et al., 2009) N/A
Anti-mAB414 Abcam Cat#ab5008
Anti-HTP-3 (Goodyer et al., 2008) N/A
Anti-SYP-1 (MacQueen et al., 2002) N/A
Anti-RAD-51 (Colaiacovo et al., 2003) N/A
Anti-biotin Abcam Cat# ab6650; RRID: AB_305623
Chemicals, Peptides, and Recombinant Proteins
SYTO-12 ThermoFischer Scientific S7574
Vectashield Mounting Medium Vector Labs Cat#H-1000
(-)-Tetramisole hydrochloride Sigma-Aldrich Cat#L9756
Critical Commercial Assays
Gateway BP Clonase II Enzyme Mix ThermoFischer Scientific Cat#11789020
Gateway LR Clonase II Enzyme Mix ThermoFischer Scientific Cat#11791100
DIG (Digoxigenin)-nick translation Kit Sigma-Aldrich Cat#11745816910
Experimental Models: Organisms/Strains
C. elegans: N2 Bristol CGC https://cgc.umn.edu/strain/search
C. elegans: [gfp::lmn-1] CrispR: lmn-1(jf98[gfp::lmn-1]) I This paper UV117
C. elegans: [gfp::lmn-1]; plk-2(K65M): plk-2(tm1395) lmn-1(jf98[gfp::lmn-1]) I; vvSi18 [pie-1p::mCherry::plk-2(K65M), unc-119(+)] II This paper; plk-2(K65M) was obtained from Monique Zetka, McGill University, Canada UV118
C. elegans: [gfp::lmn-1]; chk-2(me64): lmn-1(jf98[gfp::lmn-1]) I; chk-2(me64) rol-9(sc148)/unc-51(e369) rol-9(sc148) V This paper UV119
C. elegans: [gfp::lmn-1] MosSCI: lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II This paper UV120
C. elegans: [gfp::lmn-1S32,403A] MosSCI: lmn-1(tm1502) I; jfSi72[Plmn-1S(32, 403)A::gfp cb-unc-119(+)] II This paper UV121
C. elegans: [gfp::lmn-1S8A] MosSCI: lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II This paper UV122
C. elegans: ieDF2: ieDF2/mIs11 (Harper et al., 2011) ieDF2
C. elegans: spo-11(ok79): spo-11(ok79) IV/nT1 [qIs51] (IV;V) (Dernburg et al., 1998) UV123
C. elegans: [gfp::lmn-1]; [sun-1::mRuby]: lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; ieSi21 [sun-1p::sun-1::mRuby::sun-1 3′UTR + Cbr-unc-119(+)] IV This paper UV124
C. elegans: [gfp::lmn-1S8A]; [sun-1::mRuby]: lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; ieSi21 [sun-1p::sun-1::mRuby::sun-1 3′UTR + Cbr-unc-119(+)] IV This paper UV125
C. elegans: [gfp::lmn-1]; [him-8::mCherry]; sun-1(jf18): lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; jfSi62 [pie-1p::mCherry::him-8::unc-543′UTR cb-unc-119 (+)], him-8(tm611) IV; sun-1(jf18) V/nT1 [qIs51] (IV;V) This paper UV126
C. elegans: [gfp::lmn-1S32,403A]; [him-8::mCherry]; sun-1(jf18): lmn-1(tm1502) I; jfSi72[Plmn-1S(32, 403)A::gfp cb-unc-119(+)] II; jfSi62 [pie-1p::mCherry::him-8::unc-543′UTR cb-unc-119 (+)], him-8(tm611) IV; sun-1(jf18) V/nT1 [qIs51] (IV;V) This paper UV127
C. elegans: [gfp::lmn-1S8A]; [him-8::mCherry]; sun-1(jf18): lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; jfSi62 [pie-1p::mCherry::him-8::unc-543′UTR cb-unc-119 (+)], him-8(tm611) IV; sun-1(jf18) V/nT1 [qIs51] (IV;V) This paper UV128
C. elegans: ced-3; [gfp::lmn-1]: lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; ced-3(n717) IV This paper UV129
C. elegans: ced-3; [gfp::lmn-1S32,403A]: lmn-1(tm1502) I; jfSi72[Plmn-1S(32, 403)A::gfp cb-unc-119(+)] II; ced-3(n717) IV This paper UV130
C. elegans: ced-3; [gfp::lmn-1S8A]: lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; ced-3(n717) IV This paper UV131
C. elegans: [gfp::lmn-1]; spo-11(ok79): lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; spo-11(ok79) IV/nT1 [qIs51] (IV;V) This paper UV132
C. elegans: [gfp::lmn-1S8A]; spo-11(ok79): lmn-1(tm1502) I; jfSi89[Plmn1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; spo-11(ok79) IV/nT1 [qIs51] (IV;V) This paper UV133
C. elegans: [sun-1S10A::gfp]∗: jfSi49[Psun-1::gfp S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)] IV; sun-1 (ok1282) V
∗This strain contains the mutated phospho sites described in (Woglar et al., 2013) and three sites found with more sensitive mass spectrometry detection (16, 69, 78).		 This paper UV134
C. elegans: [gfp::lmn-1]; [sun-1::gfp]: lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; jfSi26[Psun-1::GFP cb-unc-119(+)] IV; sun-1(ok1282) V This paper UV135
C. elegans: [gfp::lmn-1S32,403A];[sun-1S10A::gfp]: lmn-1(tm1502) I; jfSi72[Plmn-1S(32, 403)A::gfp cb-unc-119(+)] II; jfSi49[Psun-1::gfp S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)] IV; sun-1 (ok1282) V This paper UV136
C. elegans: [gfp::lmn-1S8A]; [sun-1S10A::gfp]: lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; jfSi49[Psun-1::gfp S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)] IV; sun-1 (ok1282) V This paper UV137
C. elegans: [gfp::lmn-1]; [sun-1S10A::gfp]: lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; jfSi49[Psun-1::GFP S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)] IV; sun-1 (ok1282) V This paper UV138
C. elegans: [lmn-1S32,403,470A]: lmn-1(tm1502) I; jfSi76[Plmn-1S(32,403,470)A cb-unc-119(+)] II This paper UV139
C. elegans: [gfp::lmn-1]; [sun-1::gfp] spo-11(ok79): lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; jfSi26[Psun-1::gfp cb-unc-119(+)] spo-11(ok79) IV/nT1 [qIs51] (IV;V); sun-1(ok1282) V/nT1 [qIs51] (IV;V) This paper UV140
C. elegans: [gfp::lmn-1S8A]; [sun-1S10A::gfp] spo-11(ok79): lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; jfSi49[Psun-1::gfp S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)]spo-11(ok79) IV/nT1 [qIs51] (IV;V); sun-1(ok1282) V/nT1 [qIs51] (IV;V) This paper UV141
C. elegans: [gfp::lmn-1]; histone::mCherry: lmn-1(tm1502)I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; histone::mCherry IV This paper UV142
C. elegans: [gfp::lmn-1S32,403A]; histone::mCherry: lmn-1(tm1502)I; jfSi72[Plmn-1S(32, 403)A::gfp cb-unc-119(+)] II; histone::mCherry IV This paper UV143
C. elegans: [gfp::lmn-1S8A]; histone::mCherry: lmn-1(tm1502)I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::gfp cb-unc-119(+)] II; histone::mCherry This paper UV144
C. elegans: [gfp::lmn-1]; [sun-1::gfp] ced-3(n717): lmn-1(tm1502) I; jfSi68[Plmn-1::gfp cb-unc-119(+)] II; jfSi26[Psun-1::gfp cb-unc-119(+)] ced-3(n717) IV; sun-1(ok1282) V This paper UV147
C. elegans: [gfp::lmn-1S8A]; [sun-1S10A::gfp] ced-3(n717): lmn-1(tm1502) I; jfSi89[Plmn-1S(21,22,24,32,397,398,403,405)A::GFPcb-unc-119(+)] II; jfSi49[Psun-1::gfp S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)] ced-3(n717) IV; sun-1 (ok1282) V This paper UV148
C. elegans: [sun-1S10A::gfp] ced-3(n717): jfSi49[Psun-1::gfp S(8, 12, 16, 24, 36, 43, 58, 62, 69, 78)A cb-unc-119(+)] ced-3(n717) IV; sun-1 (ok1282) V This paper UV146
Oligonucleotides
For primers see Table S3 N/A N/A
Recombinant DNA
Destination vector pDESTttTi5605[R4-R3] for MOS insertion on Chromosome II (was a gift from Erik Jorgensen) (Frøkjaer-Jensen et al., 2008) pCFJ150
Addgene plasmid # 19329
Destination vector pCFJ178 - cxTi10882_MCS
for cxTi10882 targetting region for MOS insertion on Chromosome IV (was a gift from Erik Jorgensen)		 (Frøkjaer-Jensen et al., 2008) pCFJ178
Addgene plasmid # 19331
Co-injection marker Pmyo-2::mCherry::unc-54utr (was a gift from Erik Jorgensen) (Frøkjaer-Jensen et al., 2008) pCFJ90
Addgene plasmid # 19327
Co-injection marker pGH8 - pRAB-3::mCherry::unc-54utr (was a gift from Erik Jorgensen) (Frøkjaer-Jensen et al., 2008) pGH8
Addgene plasmid # 19359
MOS transposase Pglh-2::MosTase::glh-2utr (was a gift from Erik Jorgensen) Frøkjaer-Jensen et al., 2008) pJL43.1
Addgene plasmid # 19332
Entry clone for Gateway pDONR™221 (lmn-1 CDS was cloned in this vector- also versions with point mutations) Multistite Gateway ® Kit, Thermo Fischer Scientific Cat# 12537-023
pDONR™ P4-P1R (the lmn-1 5’ UTR with and without GFP was cloned in this vector) Multistite Gateway ® Kit, Thermo Fischer Scientific Cat# 12537-023
pDONRTM P2r-P3 (the lmn-1 3’UTR was cloned in this vector) Multistite Gateway ® Kit, Thermo Fischer Scientific Cat# 12537-023
Minigene from IDT for generating the lmn-18A containing the whole CDS of lmn-1(S21,22,24,32,397,398,403,405A) This paper N/A
Plmn-1_gfp::lmn-1_lmn-1 3’UTR in pCFJ150 This paper N/A
Plmn-1_gfp::lmn-1(S8A)_lmn-1 3’UTR in pCFJ150 This paper N/A
Plmn-1_gfp::lmn-1(S32,403A)_lmn-1 3’UTR in pCFJ150 This paper N/A
Plmn-1_lmn-1(S32,403,470)_lmn-1 3’UTR in pCFJ150 This paper N/A
Plmn-1_lmn-1_lmn-1 3’UTR in pCFJ150 This paper N/A
Minigene from IDT for generating the SUN-1(10A) containing part of the CDS of SUN-1(8,12,16,24,36,43,58,62,69,78A) This paper N/A
Psun-1::sun-1::eGFP::sun-1 3’UTR Penkner et al., 2009 N/A
Peft-3::cas9-SV40_NLS::tbb-2 3'UTR was a gift from John Calarco (Friedland et al., 2013) Addgene plasmid # 46168
pRB1017 was a gift from Andrew Fire (Arribere et al., 2014) Addgene plasmid # 59936
Software and Algorithms
ImageJ version 2.0.0
Plugins used: StackReg; Manual Tracking		 NIH https://imagej.nih.gov/ij/
SoftWorx Suite Applied Precision N/A
GraphPad Prism 6 GraphPad N/A
Adobe Photoshop CC2015 Adobe N/A
Adobe Illustrator CS6 Adobe N/A
MaxQuant Software 1.5.6.0 MaxQuant www.coxdocs.org
GPS SUMO SP 2.0 The CUCKOO Workgroup http://sumosp.biocuckoo.org/online.php