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. 2021 Mar 18;81(6):1246–1259.e8. doi: 10.1016/j.molcel.2021.01.005
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies

Mouse anti-FLAG (HRP conjugate) Sigma Cat#A8592; RRID:AB_439702
Rabbit anti-INTS4 Abcam Cat#ab75253; RRID:AB_1280962
Rabbit anti-INTS11 Cusabio Cat#PA722574ESR1HU; RRID:AB_2888982
Rabbit anti-GAPDH ProteinTech Cat#10494-1-AP; RRID:AB_2263076
Mouse anti-SBP EMD Millipore Cat#MAB10764; RRID:AB_10631872
Goat anti-Rabbit Abcam Cat#ab205718; RRID:AB_2819160
Mouse anti-HA (HRP-Conjugate) Santa Cruz Cat#SC7392HRP; RRID:AB_627809
Goat anti-Mouse Thermo Fisher Cat#31430; RRID:AB_228307

Bacterial and virus strains

NEB10 NEB Cat# C3020K
DH10EMBacY Bieniossek et al., 2012 N/A

Chemicals, peptides, and recombinant proteins

RNAiMAX Invitrogen Cat#13778030
L-Glutamine GIBCO Cat#A29168-01
Penicillin-Streptomycin (10,000 U/mL) GIBCO Cat#15140-122
DMEM GIBCO Cat#31966-021
ExpressFive™ SFM GIBCO Cat#10486025
SF-900™ II SFM GIBCO Cat#11497013
Opti-MEM GIBCO Cat#31985-062
Trypsin-EDTA GIBCO Cat#25200-056
FBS GIBCO Cat#10270-106
LipoD293 Sinagen Cat#SL100668
PEI 25k Polysciences Cat#23966-1
Tween20 Sigma Cat#P1379-500ML
Freestyle™ 293 Expression Medium GIBCO Cat#12338018
Glutaraldehyde Sigma Aldrich Cat#G5882-10X1ML
Triton X-100 ICN Biomedicals Cat#807426
Pierce™ ECL Western Blotting Substrate Thermo Fisher Cat#32209
Immobilon Membrane Merck Cat#PVH000010
Anti-FLAG Resin Sigma Aldrich Cat#F2426-1ML
Anti-HA Resin Sigma Aldrich Cat#A2905-1ML
High Capacity Streptavidin Agarose Resin Thermo Fisher Cat#20361
His60 Ni Superflow Resin Takara Cat#635660
IgG Sepharose™ 6 Fast Flow GE Healthcare Cat#17-0969-01
Amylose Resin NEB Cat#E8021S

Deposited data

High-resolution cryo-EM map This paper EMDB: EMD-12165
Medium-resolution cryo-EM map This paper EMDB: EMD-12166
INTS4-focused cryo-EM map This paper EMDB: EMD-12164
CTD2-focused cryo-EM map This paper EMDB: EMD-12163
Model refined against high-resolution map This paper PDB: 7BFP
Pseudoatomic model fitted into medium-resolution map This paper PDB: 7BFQ

Experimental models: cell lines

HEK293T ATCC Cat#CRL-3216
FreeStyle™ 293-F GIBCO Cat#R79007
SF21 GIBCO Cat#11497013
Trichoplusia ni High Five Cells Invitrogen Cat#B85502

Oligonucleotides

INTS4 siRNA (5′-GUAGGCUUAAGGAGUAUGUGAUU-3′) Dharmacon, based on Albrecht et al. (2018) N/A
INTS11 siRNA (5′-CAGACUUCCUGGACUGUGUUU-3′) Dharmacon, based on Albrecht et al. (2018) N/A
Scrambled siRNA control (5′-UGCACCGAGUGGCGACACCUU-3′) this study N/A

Recombinant DNA

pBIG1a+8xHis-INTS4/INTS9/SBP-INTS11 this study N/A
pBIG1a+8xHis-INTS5/SBP-INTS8 this study N/A
pBIG1c+8xHis-INTS10/INTS13/SBP-INTS14 this study N/A
pMG-3xFLAG+INTS11 this study N/A
pMG-3xFLAG+INTS11ΔCTD2 this study N/A
pMG-3xFLAG+INTS11ΔCTD1ΔCTD2 this study N/A
pMG-3xFLAG+INTS11ΔCTD1 this study N/A
pMG_3xFLAG+INTS11H480P/T482P this study N/A
pMG-3xFLAG+INTS11V454P/G455P this study N/A
pMG-3xFLAG+INTS11F490P/L492P this study N/A
pMG-3xFLAG+INTS11K462E this study N/A
pMG-3xFLAG+INTS11R510P/T512P based on Wu et al. (2017) N/A
pMG-3xFLAG+INTS11E203Q based on Baillat et al. (2005) N/A
pMG-SBP+INTS4K122A/H124A/Q125A this study N/A
pMG-SBP+INTS4H164A/R167A this study N/A
pMG-SBP+INTS4R210A this study N/A

Software and algorithms

PyMol N/A https://pymol.org/2/
Coot Emsley and Cowtan, 2004 https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
CCPEM Burnley et al., 2017 https://www.ccpem.ac.uk/
Relion Scheres, 2012 https://www3.mrc-lmb.cam.ac.uk/relion/
UCSF ChimeraX Goddard et al., 2018 https://www.cgl.ucsf.edu/chimerax/
Image Lab Bio-Rad http://www.bio-rad.com/en-fr/product/image-lab-software?ID=KRE6P5E8Z
Fiji https://imagej.net/Fiji
MARS BMG Labtech https://www.bmglabtech.com/de/mars-datenanalyse-software/
Promals3D Pei et al., 2008 http://prodata.swmed.edu/promals3d/promals3d.php

Other

UltrAuFoil R1.2/1.3 300 mesh Quantifoil Cat#Q350AR13A
Superose 6 3.2/300 Increase GE Healthcare Cat#GE29-0915-98
Vitribot Mark IV Thermo Fisher Scientific https://www.thermofisher.com/us/en/home.html
Amicon-Ultra 0.5 ml Centrifugal filters – 50 kDa Merck UFC5050
96-well plate, black Corning Cat#3925
CLARIOstar BMG Labtech https://www.bmglabtech.com/clariostar-plus/
ChemiDoc MP Bio-Rad http://www.bio-rad.com/en-us/category/chemidoc-imaging-systems?ID=NINJ0Z15
Vibra Cell VCX750 Sonics https://www.sonics.com/liquid-processing/products/vibra-cell-processors/vcx-500-vcx-750/