REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
ATP2A2	Thermo Fisher Scientific	Cat# MA3-919; RRID:AB_325502
Calreticulin	Cell Signaling	Cat# 12238; RRID:AB_2688013
FLAG	Sigma-Aldrich	Cat# F1804; RRID: AB_262044
β-actin	Cell Signaling	Cat# 8457; RRID:AB_10950489
C4ORF3	This paper	N/A
Anti-DYKDDDDK magnetic beads	Sigma-Aldrich	Cat# M8823; RRID:AB_2637089
Streptavidin magnetic beads	Thermo Fisher Scientific	Cat# 65001
Total oxphos cocktail	Abcam	Cat# ab110413; RRID: AB_2629281
TNAP	R&D Systems	Cat# AF2910-SP
COX IV	Cell Signaling	Cat# 38563; RRID:AB_2799136
UCP1	Abcam	Cat# ab10983; RRID:AB_2241462
Goat anti-mouse IgG (H&L) secondary antibody, HRP	Thermo Fisher Scientific	Cat# 31430; RRID: AB_10960845
Goat anti-rabbit IgG (H&L) secondary antibody, HRP	Abcam	Cat# ab6721; RRID: AB_955447
Biological samples
Human adipose tissue	This paper	N/A
Chemicals, peptides, and recombinant proteins
Indomethacin	Sigma-Aldrich	Cat# I7378
Insulin	Sigma-Aldrich	Cat# I6634
Adenylyl imidodiphosphate	MedChemExpress	Cat# HY-130777
Isobutylmethylxanthine (IBMX)	Sigma-Aldrich	Cat# I5879
Dexamethasone	Sigma-Aldrich	Cat# D4902
3,3′,5-triiodo-L-thyronine (T3)	Sigma-Aldrich	Cat# T2877
L-(-)-Norepinephrine(+)-bitartrate salt monohydrate	Sigma-Aldrich	Cat# A9512
DMEM	Gibco	Cat# 11965092
Fetal Bovine Serum	ATLANTA biologicals	Cat# S11550
Penicillin-Streptomycin	Gibco	Cat# 15140
Basticidin S HCl (10 mg/mL)	Gibco	Cat# A1113903
0.05% Trypsin	Corning	Cat# MT25052CI
cOmplete, EDTA-free Protease Inhibitor Cocktail	Roche	Cat# 11873580001
Phosphatase inhibitor cocktail 2	Sigma-Aldrich	Cat# P5726
Phosphatase inhibitor cocktail 3	Sigma-Aldrich	Cat# P0044
Dextrose	Sigma-Aldrich	Cat# D9434
Palmitoyl-L-carnitine	Sigma-Aldrich	Cat# 61251
Bovine Serum Albumin	Sigma-Aldrich	Cat# A1595
Malate	Sigma-Aldrich	Cat# M1000
Succinate	Sigma-Aldrich	Cat# 224731
Glutamate	Sigma-Aldrich	Cat# G1501
Rotenone	Sigma-Aldrich	Cat# R8875
N,N.N’,N’-Tetramethyl-p-phenylenediamine	Sigma-Aldrich	Cat# T7394
L-ascorbate	Sigma-Aldrich	Cat# A4034
Thapsigargin	Sigma-Aldrich	Cat# T9033
45CaCl2	PerkinElmer	Cat# NEZ013001MC
CaCl2	Sigma-Aldrich	Cat# C3306
QIAzol lysis reagent	Qiagen	Cat# 79306
TRIzol reagent	Thermo Fisher Scientific	Cat# 15596026
Adenosine 5’-diphosphate	Sigma-Aldrich	Cat# A5285
MgCl2	Sigma-Aldrich	Cat# M2393
NaCl	Sigma-Aldrich	Cat# S7653
KCl	Sigma-Aldrich	Cat# P9541
Na2HPO4	Fluka	Cat# 71639
NaH2PO4	Sigma-Aldrich	Cat# 71505
Adenosine 5’-triphosphate	Sigma-Aldrich	Cat# A2383
Sucrose	Sigma-Aldrich	Cat# S0389
D-mannitol	Sigma-Aldrich	Cat# M4125
Scintillation fluid	PerkinElmer	Cat# 6013326
Tris	Sigma-Aldrich	Cat# 11814273001
EGTA	Sigma-Aldrich	Cat# E4378
n-Dodecyl β-D-maltoside	Sigma-Aldrich	Cat# D4641
DTT	Sigma-Aldrich	Cat# 43816
Glycerol	Sigma-Aldrich	Cat# G7793
3X-FLAG peptide	Sigma-Aldrich	Cat# F4799
Paraformaldehyde	Santa Cruz Biotechnology	Cat# SC281692
PBS	Gibco	Cat# 10010023
NaN3	Sigma-Aldrich	Cat# S2002
MES potassium salt	Santa Cruz	Cat# sc-286156
Chloroform	Sigma-Aldrich	Cat# 650498
Acetonitrile	Thermo Fisher Scientific	Cat# A955
Methanol	Thermo Fisher Scientific	Cat# A456
Rosiglitazone	Enzo Life Sciences	Cat# ALX-350-125
Hygromycin B	Thermo Fisher Scientific	Cat# 10687010
2,2,2-Tribromoethanol	Sigma-Aldrich	Cat# T48402
KH2PO4	Sigma-Aldrich	Cat# P5655
GlutaMAX supplement	Thermo Fisher Scientific	Cat# 30055061
Antimycin A	Sigma-Aldrich	Cat# A8674
Water LC/MS	Thermo Fisher Scientific	Cat# W64
Formic acid	Thermo Fisher Scientific	Cat# A117
Guanosine 5’-diphosphate	Sigma-Aldrich	Cat# G7127
SDS	Thermo Fisher Scientific	Cat# BP166
HEPES	Sigma-Aldrich	Cat# H3375
LiCl	Sigma-Aldrich	Cat# L4408
EDTA	Sigma-Aldrich	Cat# E5134
NP-40	Boston Bioproducts	Cat# P-877
Sodium deoxycholate	Sigma-Aldrich	Cat# D6750
Triton-X 100	Thermo Fisher Scientific	Cat# BP151
Tween 20	Thermo Fisher Scientific	Cat# BP337
KOH	Boston Bioproducts	Cat# BZ-8038
Rac-Glycerol 1-Phosphate	Santa Cruz	Cat# sc-215789
Pyruvate	Sigma-Aldrich	Cat# P4562
HBSS	Thermo Fisher Scientific	Cat# 88284
XF calibrant solution	Agilent	Cat# 100840-000
SBI-425	MedChemExpress	Cat# HY-124756
Creatine	Sigma-Aldrich	Cat# C3630
Laemmli sample buffer	Bio-rad	Cat# 1610747
β-mercaptoethanol	Sigma-Aldrich	Cat# M3148
Potassium oxalate	Sigma-Aldrich	Cat# P0963
MOPS	Sigma-Aldrich	Cat# M3183
Lauryl maltose neopentyl glycol	Thermo Fisher Scientific	Cat# A50940
Biotin	Sigma-Aldrich	Cat# B4639
Urea	Sigma-Aldrich	Cat# 51456
Bismaleimidohexane	Thermo Fisher Scientific	Cat# 22330
Polybrene transfection reagent	Sigma-Aldrich	Cat# TR-1003-G
KF	Sigma-Aldrich	Cat# 449148
AlCl3	Sigma-Aldrich	Cat# 237051
CL 316,243	Sigma-Aldrich	Cat# C5976
Phenylmethanesulfonyl fluoride	Sigma-Aldrich	Cat# 93482
DNAse	Qiagen	Cat# 79254
ERCC RNA spike-in mix	Thermo Fisher Scientific	Cat# 4456740
Critical commercial assays
RNeasy lipid tissue mini kit	Qiagen	Cat# 74804
Fluo-8 calcium flux assay kit	Abcam	Cat# ab112129
EnzChek phosphate assay kit	Thermo Fisher Scientific	Cat# E6646
Triglyceride quantification kit	Abcam	Cat# ab65336
iScript Reverse Transcription Supermix for rt-qPCR	Bio-rad	Cat# 1708841
iTaq Universal SYBR Green Supermix	Bio-rad	Cat# 1725125
Biorad gels 4-20% 15 well	Bio-rad	Cat# 4561096
Seahorse XF24 islet capture Fluxpak	Agilent	Cat# 101122-100
Pierce BCA Protein Assay Kit	Thermo Fisher Scientific	Cat# 23225
Rat/mouse insulin ELISA	Sigma-Aldrich	Cat# EZRMI
Seahorse XFe24 FluxPak	Agilent	Cat# 102340-100
Glucometer	Abbott	Cat# Freestyle Lite
Glucose strips	Abbott	Cat# 70827
Biorad gels 12% 10 well	Bio-rad	Cat# 4568044
RNeasy mini kit	Qiagen	Cat# 74104
FastSelect –rRNA HMR Kits	Qiagen	Cat# 334386
NEBNext Ultra II RNA Library Prep Kit for Illumina	New England Biolabs	Cat# E7770L
Deposited data
RNA sequencing	This paper	GSE268770
Proteomics	This paper	PXD059431
Experimental models: Cell lines
ALN Mutant_A overexpressing 293T	This paper	N/A
ALN Mutant_B overexpressing 293T	This paper	N/A
ALN Mutant_C overexpressing 293T	This paper	N/A
ALN Mutant_TM overexpressing 293T	This paper	N/A
ALN wild type overexpressing 293T	This paper	N/A
C4orf3-TurboID immortalized inguinal SVF	This paper	N/A
CRISPRi-C4orf3 immortalized inguinal SVF	This paper	N/A
W48C ALN mutant-overexpressing inguinal SVF	This paper	N/A
Immortalized inguinal SVF	Kajimura Lab43	N/A
SERCA2b-overxpressing inguinal adipocytes	This paper	N/A
Experimental models: Organisms/strains
Mouse: C57BL6J mice	Jackson Laboratory	Cat# 000664
Mouse: CRISPRi-C4orf3	This paper	N/A
Mouse: dCas9/KRAB	Kajimura lab51	N/A
Mouse: UCP1 KO	Jackson Laboratory	Cat# 003124
Mouse: UCP1 x C4orf3 double knockout	This paper	N/A
Oligonucleotides
A full list of qPCR primers in Table S2	This paper	N/A
Recombinant DNA
AAV8-CAG-EGFP-U6-Scramble	VectorBuilder	N/A
AAV8-CAG-EGFP-U6-gRNA-long tracr	VectorBuilder	N/A
Software and algorithms
CaIR	Banks Lab	https://calrapp.org/
Biorender	Biorender	https://biorender.com/
Protter	Protter	https://wlab.ethz.ch/protter/start/
GO Enrichment Analysis	GeneOntology	http://geneontology.org/
GraphPad Prism 8	GraphPad Software	https://www.graphpad.com/scientific-software/prism/
ImageJ	ImageJ Software	https://imagej.nih.gov/ij/download.html
R	Open Source	https://www.r-project.org/
Trimmomatic v.0.36	Open Source	http://www.usadellab.org/cms/?page=trimmomatic
STAR aligner v2.5.2b	Open Source	https://github.com/alexdobin/STAR
Subread package v1.5.2	Open Source	https://subread.sourceforge.net/
edgeR	Open Source	https://bioconductor.org/packages/release/bioc/html/edgeR.html
clusterProfiler	Open Source	https://bioconductor.org/packages/release/bioc/html/clusterProfiler.html
Pheatmap	Open Source	https://www.rdocumentation.org/packages/pheatmap/versions/1.0.12/topics/pheatmap
HDOCK server	Open Source	http://hdock.phys.hust.edu.cn
AlphaFold server	Open Source	https://alphafoldserver.com
Other
Standard Diet	Lab Diet	Cat# 5008