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. Author manuscript; available in PMC: 2024 Dec 21.
Published in final edited form as: Cell. 2023 Dec 5;186(26):5812–5825.e21. doi: 10.1016/j.cell.2023.11.009

Key resources table.

REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Anti-BLVRB rabbit monoclonal Sino Biological Cat# 13151-R009
Anti-HO2 mouse monoclonal Abnova Cat# H00003163- M01; RRID:AB_425487
Anti-IRS1 rabbit polyclonal EMD Millipore Cat# 06–248, RRID:AB_2127890
Anti-INSR-beta rabbit polyclonal Santa Cruz Cat# sc-711, RRID:AB_631835)
Anti-INSR-beta phospho-Y1135,1136,1150,1151 rabbit monoclonal Invitrogen Cat# MA5–15148 RRID:AB_10982593
Anti-IRS1 phospho-Y608 rabbit polyclonal EMD Millipore Cat# 09–432, RRID:AB_1163457
Anti-NOS1 rabbit monoclonal Cell Signaling Cat# 4231 RRID:AB_2152485
Anti-NOS2 rabbit monoclonal Santa Cruz Cat# sc-8310 RRID: AB_2152867
Anti-NOS3 rabbit monoclonal Cell Signaling Cat#:9572 RRID:AB_329863
Anti-NOS3 phospho-S1177 rabbit polyclonal ThermoFisher Cat# PA5–97371, RRID:AB_2809173
Anti-NOS1 phospho-S1417 rabbit polyclonal Abcam Cat#ab5583 RRID:AB_304964
Anti-AKT(pan) rabbit monoclonal Cell Signaling Cat# 4691, RRID:AB_915783
Anti-AKT phospho-S473 rabbit polyclonal Cell Signaling Cat# 9271, RRID:AB_329825
Anti-p97 mouse monoclonal Fitzgerald Industries Cat# 10R-P104a, RRID:AB_1287614
Anti-GAPDH mouse monoclonal Abcam Cat# ab181602, RRID:AB_2630358
Anti-AS160 rabbit monoclonal ThermoFisher Cat# MA5–14840, RRID:AB_10979793
Anti-AS160 phospho-T642 rabbit polyclonal ThermoFisher 4Cat# 44–1071G, RRID:AB_2533564
Anti-ACADVL Santa Cruz Cat#sc-271225 RRID:AB_10609094
Anti-FLAG M2 Sigma-Aldrich Cat# F3165, RRID:AB_259529
Anti-AKR1A1 mouse monoclonal Origene Cat#TA500740 RRID:AB_11128818
Anti-AKR1A1 mouse monoclonal Santa Cruz Cat# sc-100500 RRID:AB_1118799
Anti-Histone3 rabbit Cell Signaling Cat#9705 RRID:AB_331563
Anti-cytochrome C mouse monoclonal Cell Signaling Cat#12963 RRID:AB_2637072
Anti-c-myc mouse monoclonal R&D system Cat#MAB3696
Anti-Flag mouse monoclonal Sigma-Aldrich Cat#F1804 RRID:AB_262044
Anti-His-Tag Cell Signaling Cat#2365 RRID:AB_2115720
Anti-c-myc agarose Sigma-Aldrich Cat# A7470, RRID:AB_10109522
Anti-FLAG M2-agarose Sigma-Aldrich Cat# A2220, RRID:AB_10063035
Bacterial and virus strains
BL21-CodonPlus E. coli Agilent Cat# 230245
Biological samples
De-identified human skeletal muscle samples, human visceral adipose samples and human subcutaneous adipose tissue samples Tissue Resource Core at University Hospitals Cleveland/CWRU https://cwru.corefacilities.org/service_center/show_external/4499?name=tissue-resource-core
Bovine liver Rockland Cat# BV-T278 Rockland Cat# BV-T278
Fetal bovine serum (FBS) Sigma-Aldrich Cat# F4135 Sigma-Aldrich Cat# F4135
Chemicals, peptides, and recombinant proteins
Coenzyme A-agarose powder Sigma-Aldrich Cat# C7013
Amylose-resin BioLabs Cat# E8021S
Activated thiol-Sepharose 4B GE Healthcare Cat# 17–0640-01
Thiopropyl-Sepharose 6B GE Healthcare Cat# 17–0420-01
Acetyl-CoA-agarose This study N/A
Palmitoyl-CoA-agarose Sigma-Aldrich Cat# P5297
Glutathione (reduced)-agarose Invitrogen Cat# G2879
SNO-modified activated thiol-Sepharose 4B This study N/A
SNO-modified coenzyme A (GSNO)-agarose This study N/A
Anti-TMT resin ThermoFisher Cat# 90060
Protein A/G-Agarose Thermo Scientific Cat# 20423
Ni-NTA-agarose Thermo Scientific Cat# 88221
3X FLAG peptide Sigma-Aldrich Cat# F4799
[3H]-2-deoxyglucose PerkinElmer Cat# NET549250UC
[14C]-mannitol PerkinElmer Cat# NEC852050UC
Insulin (human) Sigma-Aldrich Cat# I9278
PolyJet transfection reagent SignaGen Cat# SL100688
DMEM, high glucose Thermo Scientific Cat# 11965092
Penecillin/streptomycin Thermo Scientific Cat# 15140122
Puromycin Thermo Scientific Cat# A1113803
Krebs-Henseleit buffer Thermo Scientific Cat# J67795-AP
Sodium pyruvate Thermo Scientific Cat# 11360070
Mannitol Sigma-Aldrich Cat# M4125
2-deoxyglucose Sigma-Aldrich Cat# D8375
DPTA (diethylenetriaminepentaacetic acid) Cayman Chemical Cat# 33307
NaNO2 Fluka Cat# 71759
Bovine serum albumin (BSA) Sigma-Aldrich Cat# A2153
Neocuproine Sigma-Aldrich Cat# N1501
Protease inhibitor mix Roche Cat# 04693132001
Phenylmethylsulfonyl fluoride (PMSF) Sigma-Aldrich Cat# P7626
Imidazole Sigma-Aldrich Cat# I3386
2-mercaptoethanol Sigma-Aldrich Cat# M6250
S-methylmethanethiosulfonate (MMTS) Sigma-Aldrich Cat# M9386
Sodium ascorbate Sigma-Aldrich Cat# 11140
Critical commercial assays
QuikChange II Site-Directed Mutagenesis Kit Agilent Cat# 200523
Bicinchoninic acid (BCA) protein quantitation kit ThermoFisher Cat# 23225
Accu-Chek glucose test strip Roche Cat#07299702001
Ultra Sensitive Mouse Insulin ELISA kit Crystal Chem Cat# 90080
Bilirubin assay kit Sigma-Aldrich Cat# MAK126
iTRAQ Reagents Multiplex kit Sigma-Aldrich Cat# 4352135
iodoTMTsixplex Isobaric Label Reagent Set ThermoFisher Cat# 90101
Hemin Assay Kit BioVison Cat# K672
InsR Kinase Enzyme System Promega Cat#V3901
ADP-Glo™ Kinase Assay Promega Cat#V9101
Deposited data
Raw images for Western blots This study doi:10.17632/zz3ktsyjds.1
Experimental models: Cell lines
HEK293 cells ATCC Cat# PTA-4488, RRID:CVCL_0045
Rat L6 cells ATCC Cat# CRL-1458, RRID:CVCL_0385
HEK293 SCAN-KO cells This study N/A
HEK293 SCAN-KO/SCAN-WT re-expressing cells This study N/A
HEK293 SCAN-KO/SCAN-QTG/NAA-expressing cells This study N/A
HEK293 SCAN-KO/SCAN-C109/188R-expressing cells This study N/A
HEK293-SCoR-KO cells Stomberski Et al.9 N/A
L6 SCAN-KO cells This study N/A
L6 SCAN-KO/SCAN-WT re-expressing cells This study N/A
L6 SCAN-KO/SCAN-QTG/NAA-expressing cells This study N/A
L6 SACN-KO/SCAN-C109/188R-expressing cells This study N/A
L6 Insr-KO cells This study N/A
L6 Insr-KO/INSR-WT re-expressing cells This study N/A
L6 Insr-KO/INSR-C1083S-expressing cells This study N/A
Experimental models: Organisms/strains
Blvrbtm1(KOMP)Wtsi (SCAN-KO) mice Model Animal Research Center, Nanjing University RRID:MGI:5756156
B6.Cg-Lepob/J (ob/ob) mice Jax.org Cat# 000632, RRID:IMSR JAX:00 0632
Oligonucleotides
SCAN-KO genotyping primer Blvrb-FRT-tF1 5’-AGAGTTTGGGTCCTCCCTTTCCT-3’ ThermoFisher N/A
SCAN-KO genotyping primer Common-En2-R 5’-CCAACTGACCTTGGGCAAGAACAT-3’ ThermoFisher N/A
rat sgRNA 1 (Blvrb/SCAN) 5’-CCCCTCTGACGGTAACCTGC-3’ GenScript N/A
rat sgRNA 2 (Blvrb/SCAN) 5’-TCGGTGCCACCGGAAGGACC-3’ GenScript N/A
rat sgRNA 1 (Insr) 5’-TATCGACTGGTCCCGCATCCTGG-3’ GenScript N/A
rat sgRNA 2 (Insr) 5’-GCCTGATTATCAACATCCGAGGG-3’ GenScript N/A
Mutagenesis primers (See Table S5) GenScript See Table S5
Recombinant DNA
pcDNA3 human IRS1-FLAG GenScript Cat# OHu24973
pcDNA3 human INSR-FLAG Genscript Cat# OHu24951
pcDNA3.1 vector ThermoFisher Cat# V79020
pcDNA3.1 human HO2 This study N/A
pcDNA3.1 human HO2 C127R mutant This study N/A
pcDNA3.1 human HO2 C265R mutant This study N/A
pcDNA3.1 human HO2 C282R mutant This study N/A
pcDNA3.1 human HO2 1–296 This study N/A
pcDNA3.1 human HO2 65–316 This study N/A
pcDNA3.1 human HO2 130–316 This study N/A
pcDNA3.1 human HO2 195–316 This study N/A
pcDNA3.1 human SCAN(BLVRB) This study N/A
pcDNA3.1 human SCAN QTG-NAA mutant This study N/A
pcDNA3.1 human SCAN C109R mutant This study N/A
pcDNA3.1 human SCAN C188R mutant This study N/A
pcDNA3.1 human SCAN C109/188R mutant This study N/A
pcDNA3 human INSR C825A mutant This study N/A
pcDNA3 human INSR C834A mutant This study N/A
pcDNA3 human INSR C1083A mutant This study N/A
pcDNA3 human NOS3 (eNOS) Ozawa Et al. 70 N/A
pET21b vector Sigma-Aldrich Cat# 69741
pET21b human HO2–6xHis This study N/A
pET21b human HO2–6xHis-195–316 This study N/A
pET21b human SCAN-6xHis This study N/A
pET21b human SCAN-6xHis QTG-NAA mutant This study N/A
pET21b human SCAN-6xHis C109/188R mutant This study N/A
Human BLVRB CRISPR/Cas9 KO plasmid Santa Cruz Biotechnology Cat# sc-405988
Human BLVRB HDR plasmid Santa Cruz Biotechnology Cat# sc-405988- HDR
Rat eSpCas9-Blvrb targeting plasmid GenScript N/A
Rat eSpCas9-Insr targeting plasmid GenScript N/A
Software and algorithms
GraphPad Prism, v8 GraphPad Inc https://www.graphpad.com/
Other
High-fat diet for mice (60 kcal% fat diet) Research Diets, Inc. D12492
Teklad rodent chow diet Envigo P3000