REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit polyclonal anti-Drp1 (1:1000)	This paper	N/A
Rabbit polyclonal anti-phospho Drp1 Ser600 (1:2000)	This paper	N/A
Rabbit polyclonal anti-phospho Drp1 Ser579 (1:1000)	This paper	N/A
Rabbit monoclonal antibody anti-GAPDH (Clone 14C10) (1:2000)	Cell Signaling Technology	Cat#2118; RRID: AB_561053
Rabbit polyclonal anti-Pan-cadherin (1:1000)	Cell Signaling Technology	Cat#4068; RRID: AB_2158565
Rabbit monoclonal anti- Rb (D20) (1:1000)	Cell Signaling Technology	Cat#9313; RRID: AB_1904119
Rabbit polyclonal phospho-Rb (Ser780) (1:1000)	Cell Signaling Technology	Cat# 9307; RRID: AB_330015
Rabbit monoclonal anti-Cdk1 (1:1000)	Abcam	Cat#ab133327; RRID: AB_11155333
Rabbit polyclonal anti-phospho-Cdk1 (Tyr15) (1:500)	Millipore	Cat# 219440; RRID: AB_564425
Rabbit polyclonal anti-Mfn1(1:2000)	Kulkarni et al., 2016	N/A
Rabbit polyclonal anti-Mfn2 (1:2000)	Kulkarni et al., 2016	N/A
Mouse monoclonal anti- OPA1 (Clone 18/OPA1) (1:2000)	BD Biosciences	Cat# 612606; RRID: AB_399888
Mouse monoclonal anti- Anti-VDAC1 / Porin (1:5000)	Abcam	Cat#ab14734; RRID: AB_443084
Mouse monoclonal anti-NDUFA9 (Clone 20C11) (1:2000)	Abcam	Cat# ab14713; RRID: AB_301431
Mouse monoclonal anti-SDHA (Clone 2E3) (1:5000)	Abcam	Cat# ab14715; RRID: AB_301433
Mouse monoclonal anti-UQCRC1 (1:2000)	Abcam	Cat# ab110252; RRID: AB_10863633
Mouse monoclonal anti-MTCO1 (1:1000)	Abcam	Cat#ab14705; RRID: AB_2084810
Mouse monoclonal anti-ATP5A (1:5000)	Abcam	Cat# ab14748; RRID: AB_301447
Rabbit polyclonal anti-UCP1 (1:5000)	Abcam	Cat# ab10983; RRID: AB_2241462
Bacterial and virus strains
DH5alpha competent	ThermoFisher	#18265017
Chemicals, peptides, and recombinant proteins
Oligomycin	Sigma	#1404-19-9
FCCP	Sigma	#370-86-5
Rotenone	Sigma	#83-79-4
Antimycin A	Sigma	#1397-94-0
CL 316,243	Sigma	#C5976
Glucagon	Novo Nordisk	#0169-7065-15
Isoproterenol hydrochloride	Sigma	#I6504
Glucose	GIBCO	#15023021
Insulin (100U/mL)	NovoRapid	# 058610
Roscovitine	Cell Signaling Technology	#9885
KOD Hot Start DNA Polymerase	Merck/SigmaAldrich	71086
T4 DNA ligase	Fermentas/Thermofisher	#EL0016
BamH1	Fermentas/Thermofisher	#FD0055
Not1	Fermentas/Thermofisher	#ER0592
Dpn1	Fermentas/Thermofisher	#ER1702
Critical commercial assays
Proteome Profiler Mouse Adipokine Array Kit	R&D Systems	#ARY013
DNeasy Blood & Tissue Kit	QIAGEN	#69504
BigDye Terminator 3.1 kit	Applied Biosystems	# 4337454
NucleoSpin® Plasmid	MACHEREY-NAGEL	740499
NucleoSpin Gel and PCR clean-up kit	MACHEREY-NAGEL	740609
Deposited data
Metabolomics	MetaboLight	MTBLS3035
Proteomics	ProteomeXchange	PXD027108
Experimental models: Cell lines
Mouse: Drp1 KO mouse embryonic fibroblasts	Ishihara et al., 2009	N/A
Mouse: Drp1 WT mouse embryonic fibroblasts	Ishihara et al., 2009	N/A
Experimental models: Organisms/strains
Mouse: Dnm1l S643A knock-in (KI)	Taconic	C57BL/6NTac-Dnm1ltm3143(S643A)Arte
Mouse: Dnm1l S643A wild type (WT)	Taconic	C57BL/6NTac
Oligonucleotides
Primers for cloning, see Table S1	This paper	N/A
Primers for RT-qPCR, see Table S2	This paper	N/A
Recombinant DNA
Plasmid: pcDNA3.1 (+)	Thermofisher	V79520
Plasmid: pcDNA3.1(+) Drp1	A gift from Dr. David Chan, California Institute of Technology (Caltech, USA)	Addgene reference #34706
Plasmid: pcDNA3.1 Drp1 WT	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S579A	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S579E	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S579D	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S600A	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S600E	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S600D	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S579E/S600E	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S579A/S600E	This paper	N/A
Plasmid: pcDNA3.1 Drp1 S579E/S600A	This paper	N/A
Plasmid: pVITRO2-neo-mcs	InvivoGen	pvitro2-nmcs
Plasmid: pVitro2- Drp1 WT - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S579A - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S579E - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S600A - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S600E - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S579E/S600E - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S579A/S600E - mtDSRED	This paper	N/A
Plasmid: pVitro2- Drp1 S579E/S600A - mtDSRED	This paper	N/A
Software and algorithms
ImageJ	Schneider et al., 2012	https://imagej.nih.gov/ij/
xcms	Tautenhahn et al., 2012	https://xcmsonline.scripps.edu
MiNa	Valente et al., 2017	https://github.com/StuartLab/MiNA
GraphPad Prism 6	GraphPad Software	https://www.graphpad.com/