| Antibodies |
|
|
| MARCH8 polyclonal antibody (A01) |
Abnova |
Cat#H00220972-A01; RRID:AB_463085 |
| Anti-Hepatitis C Virus Core 1b antibody (C7–50) |
Abcam |
Cat#ab2740 |
| Monoclonal anti-FLAG M2 antibody |
Sigma-Aldrich |
Cat#F3165; RRID:AB_259529 |
| Mouse Ub antibody (P4D1) |
Santa Cruz Biotechnology |
Cat#SC-8017; RRID:AB_628423 |
| Anti GLuc antibody |
New England BioLabs |
Cat#E8023 |
| Monoclonal Anti-HA (HA-7) antibody |
Santa Cruz Biotechnology |
Cat#H3663 |
| Normal mouse IgG |
Santa Cruz Biotechnology |
Cat#SC-2025; RRID:AB_737182 |
| Normal Rabbit IgG Polyclonal Antibody control |
Millipore, Sigma |
Cat#12–370 |
| Anti-mouse IgG, HRP-linked Antibody |
Cell Signaling |
Cat#7076S; RRID:AB_330924 |
| Anti-Rabbit IgG HRP-linked |
Cell Signaling |
Cat#7074s |
| Goat anti-Rabbit IgG (H+L), Alexa Fluor 488 |
Invitrogen |
Cat#A27034 |
| Donkey anti-Mouse IgG (H+L) Alexa Fluor 568 |
Invitrogen |
Cat#A10037; RRID:AB_2534013 |
| Rabbit monoclonal anti-NS2 |
Gift from Dr. Brett Lindenbach |
Barouch-Bentov et al., 2016 |
| CellLight ER-GFP, BacMam 2.0 |
Invitrogen |
Cat#C10590
|
| Bacterial and virus strains |
|
|
| pFL-J6/JFH(p7-Rluc2A), Renilla reporter HCV |
Gift from Dr. Charles Rice |
Murray et al., 2007 |
| pDENV2 (New Guinea C strain), Renilla reporter |
Gift from Dr. Pei-Yong Shi |
Xie et al., 2013; Zou et al., 2011
|
| pZIKV (FSS13025, Asian strain) Renilla reporter |
Gift from Dr. Pei-Yong Shi |
Shan et al., 2016 |
| Chemicals, Peptides, and Recombinant Proteins |
|
|
| PR619 |
LifeSensors |
Cat#SI9619 |
| 1, 10- phenanthroline |
LifeSensors |
Cat#SI9649 |
| Dithiobis-succinimidyl-propionate (DSP) |
Pierce |
Cat#22586 |
| Protease Inhibitor Cocktail |
Sigma-Aldrich |
Cat#P8340 |
| N-Ethylmaleimide (NEM) |
Sigma-Aldrich |
Cat#E3876 |
| Lipofectamine 3000 |
Life Technologies |
Cat#L3000–001 |
| Cycloheximide (CHX) |
Sigma-Aldrich |
Cat#C7698 |
| siIMPORTER |
Millipore |
Cat# 64–101 |
| Dynabeads protein A/G |
Life technology |
Cat#10002d; Cat# 10004d |
| AlamarBlue |
Invitrogen |
Cat#DAL1100 |
| Anti-K63 TUBEs |
LifeSensors |
Cat#UM604 |
| Recombinant MARCH8 |
Oregene |
Cat#TP305477 |
| Recombinant NS2 |
Einav lab |
Barouch-Bentov et al., 2016 |
| Critical Commercial Assays |
|
|
| Ubiquitinylation kit |
ENZO |
Cat#BML-UW9920–0001 |
| Quickchange Lightning kit |
Agilent |
Cat# 210519 |
| QuickTiter HCV Core Antigen ELISA Kit |
Cell BioLabs |
Cat#VPK-151 |
| NucleoSpin RNA Virus kit |
MACHEREY-NAGEL |
Cat# 740956.250 |
| Power SYBR Green RNA-to-CT 1-Step Kit |
Applied Biosystems |
Cat# 4389986 |
| Experimental Models: Cell Lines |
|
|
| Homo sapiens: HEK293T |
ATCC |
CRL-3216 |
| Homo sapiens: Huh7.5 |
Apath, LLC |
N/A |
| Homo sapiens: Huh7.5.1 |
Apath, LLC |
N/A |
| Homo sapiens: HEK293T MARCH8KO |
This paper |
N/A |
| Homo sapiens: Huh7.5.1 MARCH8KO#1 |
This paper |
N/A |
| Homo sapiens: Huh7.5.1 MARCH8KO#2 |
This paper |
N/A |
| Experimental Models: Organisms/Strains |
|
|
| E.coli C41 (DE3) Competent Cells |
Lucigen |
Cat#60442–1 |
| E.coli DH5α Competent Cells |
Thermofisher-Scientific |
Cat#18265017 |
| Oligonucleotides |
|
|
| ON-TARGETplus SMART pools siRNA library, see Table S5
|
GE Healthcare Dharmacon |
N/A |
| MARCH8 knockdown siRNA-1 |
Life Technologies |
Cat# s47920 |
| MARCH8 knockdown siRNA-2 |
Life Technologies |
Cat# s47921 |
| MARCH8 sgRNA-1 |
This paper |
5′- CACCGGCTCATCCCAACC TCTTATC |
| MARCH8 sgRNA-2 |
This paper |
5′- CACCGGTGCGAGAGAAGG AGGACAC |
| Primers for qPCR analysis, see Table S6
|
This paper |
N/A |
| Recombinant DNA |
|
|
| pMARCH8-LD mutant |
This paper |
van de Kooij et al., 2013 |
| pGLuc-NS2 4KE mutant |
Einav lab |
Barouch-Bentov et al., 2016 |
| pFLAG-MARCH8 |
This paper |
N/A |
| pRAB7-GFP |
Gift from Dr. Suzanne Pfeffer |
N/A |
| pHA-UB |
Gift from Dr. Brett Lindenbach |
Barouch-Bentov et al., 2016 |
| pHA-UBK48R mutant |
Gift from Dr. Brett Lindenbach |
N/A |
| pHA-UBK63R mutant |
Gift from Dr. Brett Lindenbach |
Barouch-Bentov et al., 2016 |
| pX458 gRNA |
Gift from Dr. Feng Zhang |
Addgene plasmid # 48138 |
| Unique ORFs in the UPS library, see Table S1
|
This paper |
N/A |
| Software and Algorithms |
|
|
| Prism 7 |
Graphpad |
https://www.graphpad.com/scientificsoftware/prism/ |
| CRISPR design tool |
Chopchop |
http://chopchop.cbu.uib.no/ |
| Cytoscape |
Cytoscape |
https://cytoscape.org/ |
| ImageJ |
NIH |
https://imagej.nih.gov/ij/ |
