REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit anti-ZWILCH	Andrea Musacchio, MPI of Molecular Physiology, Dortmund, Germany	N/A
Mouse anti-ZWILCH	Andrea Musacchio	N/A
Rabbit anti-ZW10	Andrea Musacchio	N/A
Rabbit anti-BUB-1	Arshad Desai, University of California San Diego, La Jolla, CA, USA	OD31
Rabbit anti-mCherry	Arshad Desai	OD78
Rabbit anti-DSN1	Arshad Desai	OD110
Rabbit anti-KNL-1	Arshad Desai	OD33
Rabbit anti-KNL1/CASC5	Arshad Desai	OD111
Rabbit anti-ROD-1	Arshad Desai	OD216
Rabbit anti-SPDL-1	Arshad Desai	OD164
Rabbit anti-SPDL1/CCDC99	Arshad Desai	OD157
Rabbit anti-SPDL1/CCDC99	Arshad Desai	OD174
Rabbit anti-ZWL-1	Arshad Desai	OD85
Sheep anti-BUB3	Stephen Taylor, University of Manchester, UK	N/A
Sheep anti-BUBR1	Stephen Taylor	N/A
Sheep anti-CENP-F	Stephen Taylor	N/A
Goat anti-GFP	Anthony Hyman, Max Planck Institute of Molecular Cell Biology & Genetics, Dresden, Germany	N/A
Mouse anti-α-Tubulin (DM1α)	Sigma-Aldrich	Cat No.: T6199; RRID: AB_477583
Mouse anti-α-Tubulin (clone B512)	Sigma-Aldrich	Cat No.: T5168; RRID: AB_477579
Mouse anti-BUB1	Abcam	Cat No.: ab54893; RRID: AB_940664
Guinea pig anti-CENP-C	MBL	Cat No.: PD030; RRID: AB_10693556
Mouse anti-CENP-E (clone1H12)	Abcam	Cat No.: ab5093; RRID: AB_304747
Human anti-centromere antibodies (ACA)	Antibodies Incorporated	Cat No.: 15-234-0001; RRID: AB_2687472
Mouse anti-FLAG M2	Sigma-Aldrich	Cat No.: F1804; RRID: AB_262044
Mouse anti-GFP (clone 9F9.F9)	Abcam	Cat No.: ab1218; RRID: AB_298911
Mouse anti-HEC1 (clone 9G3)	Abcam	Cat No.: ab3613: RRID: AB_303949
Mouse anti-MAD1 (clone BB3-8)	Millipore	Cat No.: MABE867/6B7115
Mouse anti-MAD2	Santa Cruz Biotechnology	Cat No.: sc-65492; RRID: AB_831526
Mouse anti-nuclear pore complex antibody (clone Mab414)	Abcam	Cat No.: ab24609; RRID: AB_448181
Mouse anti-p150/DCTN1	BD Transduction Laboratories	Cat No.: 610473; RRID: AB_397845
Rabbit anti-ROD/KNTC1	[15]	GC7
Donkey anti-goat IgG, Alexa 488 conjugate	Jackson ImmunoResearch	Cat No.: 705-545-147; RRID: AB_2336933
Donkey anti-guinea pig IgG, Alexa 647 conjugate	Jackson ImmunoResearch	Cat No.: 706-605-148; RRID: AB_2340476
Donkey anti-mouse IgG, Alexa 488 conjugate	Jackson ImmunoResearch	Cat No.: 715-545-150; RRID: AB_2340846
Donkey anti-mouse IgG, Alexa 594 conjugate	Jackson ImmunoResearch	Cat No.: 715-585-150; RRID: AB_2340854
Donkey anti-mouse IgG, Alexa 647 conjugate	Jackson ImmunoResearch	Cat No.: 715-605-150; RRID: AB_2340862
Donkey anti-rabbit IgG, Alexa 488 conjugate	Jackson ImmunoResearch	Cat No.: 711-545-152; RRID: AB_2313584
Donkey anti-rabbit IgG, Alexa 594 conjugate	Jackson ImmunoResearch	Cat No.: 711-585-152; RRID: AB_2340621
Donkey anti-sheep IgG, Alexa 488 conjugate	Jackson ImmunoResearch	Cat No.: 713-545-147; RRID: AB_2340745
Bovine anti-goat IgG HRP	Jackson ImmunoResearch	Cat No.: 805-035-180; RRID: AB_2340874
Goat anti-mouse IgG HRP	Jackson ImmunoResearch	Cat No.: 115-035-003; RRID: AB_10015289
Goat anti-rabbit IgG HRP	Jackson ImmunoResearch	Cat No.: 111-035-003; RRID: AB_2313567
Bacterial and Virus Strains
DH5alpha Chemically Competent E. coli	Thermo Fisher Scientific	Cat No.: 18265017
DH10EMBacY	Geneva Biotech	N/A
TOP10 Chemically Competent E. coli	Thermo Fisher Scientific	Cat No.: C404010
Chemicals, Peptides, and Recombinant Proteins
RO-3306	Sigma-Aldrich	Cat No.: SML0569
Dulbecco's modified Eagle's medium	Gibco	Cat No.: 31966021
EDTA-free Complete Protease Inhibitor Cocktail	Roche	Cat No.: 11873580001
Fetal bovine serum	Gibco	Cat No.: 10500064
FuGENE 6 transfection reagent	Promega	Cat No.: E2691
GlutaMAX	Gibco	Cat No.: 35050038
HIS-Select Nickel Affinity Gel beads	Sigma-Aldrich	Cat No.: P6611-25ML
Hygromycin B	Life Technologies	Cat No.: LTI 10687-010
IgG-free BSA	Jackson ImmunoResearch	Cat No.: 001-000-161
Nocodazole	Sigma-Aldrich	Cat No.: M1404-2MG
Oligofectamine	Invitrogen	Cat No.: 12252-011
Opti-MEM	Gibco	Cat No.: 31985047
Pierce ECL Western Blotting Substrate	Thermo Fisher Scientific	Cat No.: 32106
Prolong Gold with DAPI stain	Invitrogen	Cat No.: P36930
SFM4 medium	Hyclone	Cat No.: SH30913.02
Strep-Tactin, Oyster 645 conjugate	IBA	Cat No.: 2-1553-050
X-tremeGene HP DNA Transfection Reagent	Roche	Cat No.: 000000006366244001
Critical Commercial Assays
MEGAscript T3	Invitrogen	Cat No.: AM1338M
MEGAscript T7	Invitrogen	Cat No.: AM1334
NucleoSpin Gel and PCR Clean-up	Macherey-Nagel	Cat No.: 1197-2372
Nucleospin Plasmid Miniprep Kit	Macherey-Nagel	Cat No.: 1235-3358
Experimental Models: Cell Lines
HeLa Flp-In T-Rex	Stephen Taylor	N/A
HeLa FRT-TO MYC::EGFP::TEV::S-peptide:: ROD/KNTC1(1-2209)	[15]	N/A
HeLa FRT-TO MYC::EGFP::TEV::S-peptide:: ROD/KNTC1(376-2209)	[15]	N/A
HeLa FRT-TO MYC::EGFP::TEV::S-peptide::ZWILCH	this study	N/A
HeLa FRT-TO MYC::EGFP::TEV::S-peptide::ZWILCH(E422A/D426A)	this study	N/A
Sf21 insect cells	Thermo Fisher Scientific	Cat No.: 11497013
Experimental Models: Organisms/Strains
C. elegans strain N2: wild-type (ancestral N2 Bristol)	Caenorhabditis Genetics Center (CGC), University of Minnesota, MN, USA	WB strain: N2 (ancestral)
C. elegans strain GCP24: unc-119(ed3) III; ruIs32[pAZ132; Ppie-1::GFP::his-58; unc-119(+)] III; ddIs6[Ppie-1::GFP::tbg-1; unc-119(+)] V; ltSi426[pDC192; Prod-1::mCherry::rod-1::rod-1 3’UTR; cb-unc-119(+)] II	[15]	GCP24
C. elegans strain GCP287: prtSi103[pRG499; Prod-1::mCherry::rod-1(373-2177)::rod-1 3’UTR; cb-unc-119(+)] II; unc-119(ed3) III	this study	GCP287
C. elegans strain GCP303: prtSi103[pRG499; Prod-1::mCherry::rod-1(373-2177)::rod-1 3’UTR; cb-unc-119(+)] II; unc-119(ed3) III?; ruIs32[pAZ132; Ppie-1::GFP::his-58; cb-unc-119(+)] III; ddIs6[Ppie- 1::GFP::tbg-1; cb-unc-119(+)] V	this study	GCP303
C. elegans strain GCP529: rod-1(lt62[gfp::rod-1]) IV; ltIs122[pAA64; Ppie-1::mCherry::his-58; cb-unc-119(+)]; unc-119(ed3) III	this study	GCP529
C. elegans strain GCP539: prtSi103[pRG499; Prod-1::mCherry::rod-1(373-2177)::rod-1 3’UTR; cb-unc-119(+)] II; unc-119(ed3) III; czw1[prt78(3xflag::czw-1)] IV	this study	GCP539
C. elegans strain GCP736: prtSi141[pRG972; Prod-1::3xflag::rod-1(373-2177)::rod-1 3’UTR; cb-unc-119(+)] II; unc-119(ed3)] III	this study	GCP736
C. elegans strain OD347: ltSi4[pOD833; Phcp-3::gfp::hcp-3::hcp-3 3’UTR; cb-unc-119(+)] II; hcp-3(ok1892) III	Arshad Desai	OD347
C. elegans strain OD933: ltSi426[pDC192;Prod-1::mCherry::rod-1::rod-1 3’UTR; cb-unc-119(+)] II; unc-119(ed3) III	[15]	OD933
C. elegans strain OD3367: rod-1(lt62[gfp::rod-1]) IV	this study	OD3367
Oligonucleotides
siRNA targeting ROD/KNTC1: GUAAAUAACUUGCGAGAGU	Dharmacon	Cat No.: J-006829-05-0020
siRNA targeting SPDL1/CCDC99: GAAAGGGUCUCAAACUGAA	Dharmacon	Cat No.: LU-016970-00-0005
siRNA targeting ZWILCH: GGUAAGAUGUGACAGUUCA	Dharmacon	Cat No.: J-019377-05-0005
Pool of 4 siRNAs targeting DCTN1: CUGGAGCGCUGUAUCGUAA GAAGAUCGAGAGACAGUUA GCUCAUGCCUCGUCUCAUU CGAGCUCACUACUGACUUA	Dharmacon	Cat No.: L-012874-00-0005
Pool of 4 siRNAs targeting KNL1/CASC5: GCAUGUAUCUCUUAAGGAA GAACGUGGGUACAAGAAGA CGAGUCAGCUUUGCAGAUA GCAAAUGACAGCCAGCUAA	Dharmacon	Cat No.: L-015673-00-0005
Control siRNA targeting Luciferase: CGUACGCGGAAUACUUCGA	Dharmacon	Cat No.: D-001100-01-20
Primers used for the production of dsRNA are listed in Table S1	this study	N/A
Recombinant DNA
Plasmid pCFJ90: Pmyo-2::mCherry::unc-54utr	Addgene	Cat No.: 19327
Plasmid pCFJ104: Pmyo-3::mCherry::unc-54utr	Addgene	Cat No.: 19328
Plasmid pGH8: Prab-3::mCherry::unc-54utr	Addgene	Cat No.: 19359
Plasmid pCFJ601: Peft-3::Mos1 transposase	Addgene	Cat No.: 34874
Plasmid pRG387: pcDNA5/FRT/TO-MYC::EGFP::TEV::S-peptide::ZWILCH	this study	pRG387
Plasmid pRG395: pcDNA5/FRT/TO-MYC::EGFP::TEV::S-peptide::ZWILCH(E422A/D426A)	this study	pRG395
Plasmid pRG442: pACEBac1-zwl-1	[15]	pRG442
Plasmid pRG445 : pACEBac1-6xHis::TEV::rod-1(1-2177)	this study	pRG445
Plasmid pRG499: pCFJ151-Prod-1::mCherry::rod-1(373-2177)::rod-1utr	this study	pRG499
Plasmid pRG516: pACEBac1-6xHis::TEV::rod-1(373-2177)	this study	pRG516
Plasmid pRG705: pACEBac1-6xHis::TEV::GFP::rod-1(1-2177)	this study	pRG705
Plasmid pRG706: pACEBac1-6xHis::TEV::GFP::rod-1(373-2177)	this study	pRG706
Plasmid pRG733: pACEBac1-StrepTagII::czw-1	this study	pRG733
Plasmid pRG972: pCFJ151-Prod-1::3xflag::rod-1(373-2177)::rod-1utr	this study	pRG972
Plasmid pL4440_DEST for RNAi of T01H3.4 (perm-1) by bacterial feeding	Ahringer RNAi Collection, Source BioScience	Cat No.: DFCIp3320H0210027D
Plasmid pOG44: Flp-recombinase expression vector	Thermo Fisher Scientific	Cat No.: V600520
Software and Algorithms
Andor iQ3	Andor Technology	https://andor.oxinst.com
FIJI (Image J version 2.0.0-rc-56/1.51h)	[50]	https://fiji.sc
Prism 7.0	GraphPad Software	https://www.graphpad.com
ZEN 2.3 (blue edition)	Zeiss	https://www.zeiss.com