1. Biochemistry and Chemical Biology
  2. Cancer Biology

Cysteine dioxygenase 1 is a metabolic liability for non-small cell lung cancer

  1. Yun Pyo Kang
  2. Laura Torrente
  3. Aimee Falzone
  4. Cody M Elkins
  5. Min Liu
  6. John M Asara
  7. Christian C Dibble
  8. Gina M DeNicola  Is a corresponding author
  1. H Lee Moffitt Cancer Center and Research Institute, United States
  2. Beth Israel Deaconess Medical Center, United States
  3. Harvard Medical School, United States
https://doi.org/10.7554/eLife.45572
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Cite this article
  1. Yun Pyo Kang
  2. Laura Torrente
  3. Aimee Falzone
  4. Cody M Elkins
  5. Min Liu
  6. John M Asara
  7. Christian C Dibble
  8. Gina M DeNicola
(2019)
Cysteine dioxygenase 1 is a metabolic liability for non-small cell lung cancer
eLife 8:e45572.
https://doi.org/10.7554/eLife.45572
  2. Download BibTeX
  3. Download .RIS
9 figures, 1 table and 1 additional file

Figures

Figure 1 with 1 supplement
Download asset Open asset
Nrf2 promotes the accumulation of intracellular cysteine and sulfur-containing metabolites.

(a) Schematic of the murine Keap1R554Q allele. The Keap1R554Q allele was created by inserting a loxP-flanked wild-type Keap1 cDNA containing exons 3–5 upstream of a R554Q mutation in the endogenous …

https://doi.org/10.7554/eLife.45572.003
Figure 1—source data 1

Nrf2 promotes the accumulation of intracellular cysteine and sulfur-containing metabolites.

https://doi.org/10.7554/eLife.45572.005
Figure 1—figure supplement 1
Download asset Open asset
Cre infection does not induce taurine pathway activity in MEFs.

(a) LC-HRMS metabolomics profiling of wild-type MEFs infected with adenoviral-cre compared to MEFs infected with empty adenovirus. N = 3 replicates/group. (b) Verification of stable-isotope labeled …

https://doi.org/10.7554/eLife.45572.004
Figure 1—figure supplement 1—source data 1

Cre infection does not induce taurine pathway activity in MEFs.

https://doi.org/10.7554/eLife.45572.006
Figure 2
Download asset Open asset
Nrf2 promotes the accumulation of cysteine dioxygenase (Cdo1) to promote entry of cysteine into the taurine synthesis pathway.

(a) Western blot analysis of Cdo1 and β-Actin levels following Nrf2 stabilization in wild-type (WT) vs. homozygous Keap1R554Q/R554Q (R554Q) MEFs. (b) Real-time PCR analysis of Cdo1 and Nqo1 mRNA …

https://doi.org/10.7554/eLife.45572.007
Figure 2—source data 1

Nrf2 promotes the accumulation of cysteine dioxygenase (Cdo1) to promote entry of cysteine into the taurine synthesis pathway.

https://doi.org/10.7554/eLife.45572.008
Figure 3 with 3 supplements
Download asset Open asset
CDO1 is preferentially silenced in KEAP1 mutant NSCLC and antagonizes proliferation.

(a) CDO1 mRNA expression of normal lung (Normal) or or KEAP1 wild-type (WT) and mutant (MUT) lung adenocarcinoma patient tumor samples. Normal, N = 45. MUT, N = 39, WT, N = 189. (b) Methylation of …

https://doi.org/10.7554/eLife.45572.009
Figure 3—source data 1

CDO1 is preferentially silenced in KEAP1 mutant NSCLC and antagonizes proliferation.

https://doi.org/10.7554/eLife.45572.016
Figure 3—figure supplement 1
Download asset Open asset
CDO1 is epigenetically silenced in NSCLC.

(a) Association of CDO1 mRNA expression with NSCLC patient outcome. (b) Correlation between CDO1 promoter methylation and mRNA expression in lung adenocarcinoma patient samples from the TCGA. N = 185…

https://doi.org/10.7554/eLife.45572.010
Figure 3—figure supplement 1—source data 1

CDO1 is epigenetically silenced in NSCLC.

https://doi.org/10.7554/eLife.45572.011
Figure 3—figure supplement 2
Download asset Open asset
CDO1 protein does not correlate with mRNA expression.

(a) Correlation between CDO1 protein levels (from Figure 3d) and CDO1 mRNA expression in the same cell lines. N = 13. (b) Western blot analysis of NRF2, xCT, CDO1 and HSP90 expression in NSCLC cell …

https://doi.org/10.7554/eLife.45572.012
Figure 3—figure supplement 2—source data 1

CDO1 protein does not correlate with mRNA expression.

https://doi.org/10.7554/eLife.45572.013
Figure 3—figure supplement 3
Download asset Open asset
NRF2 promotes CDO1 expression in NSCLC.

(a) Western blot analysis of NRF2, xCT, CDO1 and β-Actin protein expression in H460 control cells, or CDO1WT-expressing H1299 and H1975 cells following treatment with 2 µM β-lapachone (βLap). Cells …

https://doi.org/10.7554/eLife.45572.014
Figure 3—figure supplement 3—source data 1

NRF2 promotes CDO1 expression in NSCLC.

https://doi.org/10.7554/eLife.45572.015
Figure 4 with 1 supplement
Download asset Open asset
CDO1 depletes cyst(e)ine, leading to its export as CSA.

(a) Schematic depicting intracellular cysteine metabolism. Following uptake of cystine via xCT, it is reduced to cysteine via the cellular antioxidant systems. Cysteine then enters glutathione …

https://doi.org/10.7554/eLife.45572.017
Figure 4—source data 1

CDO1 depletes cyst(e)ine, leading to its export as CSA.

https://doi.org/10.7554/eLife.45572.019
Figure 4—figure supplement 1
Download asset Open asset
CSA only accounts for a fraction of CDO1-dependent (CYS)2 consumption.

(a) Time-dependent western blot analysis of CDO1 and β-Actin protein expression at 0, 1, 2, 4, 8, or 24 hr following media replenishment of CDO1WT-expressing and CDO1Y157F-expressing A549 cells. (b) …

https://doi.org/10.7554/eLife.45572.018
Figure 4—figure supplement 1—source data 1

CSA only accounts for a fraction of CDO1-dependent (CYS)2 consumption.

https://doi.org/10.7554/eLife.45572.020
Figure 5 with 4 supplements
Download asset Open asset
CDO1 expressing cells produce SO32-, which further depletes (CYS)2 via sulfitolysis.

(a) Relative abundance of cysteine-related metabolites in cell extracts following CDO1WT or CDO1Y157F expression in A549 cells. Fresh medium was added 4 hr prior to extraction. Metabolites were …

https://doi.org/10.7554/eLife.45572.021
Figure 5—source data 1

CDO1 expressing cells produce SO32-, which further depletes (CYS)2 via sulfitolysis.

https://doi.org/10.7554/eLife.45572.030
Figure 5—figure supplement 1
Download asset Open asset
Sulfite reacts with cystine and oxidized glutathione in the absence of cells.

(a) Verification of the stable-isotope labeled CYS-SO3- internal standard for absolute quantification. Stable isotope labeled CYS-SO3- was synthesized from [13C3, 15N]-cysteine and analyzed by …

https://doi.org/10.7554/eLife.45572.022
Figure 5—figure supplement 1—source data 1

Sulfite reacts with cystine and oxidized glutathione in the absence of cells.

https://doi.org/10.7554/eLife.45572.023
Figure 5—figure supplement 2
Download asset Open asset
MEFs preferentially use the CSA decarboxylation pathway.

(a) Western blot analysis of NRF2, CDO1, GOT1, CSAD and β-ACTIN levels following expression of GFP, CDO1WT, or CDO1Y157F in Keap1WT/WT (WT) and Keap1R554Q/R554Q (R554Q) MEFs. A549s expressing CDO1WT …

https://doi.org/10.7554/eLife.45572.024
Figure 5—figure supplement 2—source data 1

MEFs preferentially use the CSA decarboxylation pathway.

https://doi.org/10.7554/eLife.45572.025
Figure 5—figure supplement 3
Download asset Open asset
NRF2 promotes the CDO1-dependent production of sulfite in NSCLC cell lines.

(a) Western blot analysis of GOT1, CSAD and HSP90 levels in NRF2LOW and NRF2HIGH NSCLC cell lines. Mouse liver is included as a positive control for GOT1 and CSAD expression. Cells were fed with …

https://doi.org/10.7554/eLife.45572.026
Figure 5—figure supplement 3—source data 1

NRF2 promotes the CDO1-dependent production of sulfite in NSCLC cell lines.

https://doi.org/10.7554/eLife.45572.027
Figure 5—figure supplement 4
Download asset Open asset
CSA and sulfite are toxic to NSCLC cells.

(a) Analysis of cell viability following treatment of A549 cells with 0–10 mM HTAU, CSA or Na2SO3 for 5 days. Viable cells were analyzed with CellTiter-Glo and normalized to untreated cells. N = 3 …

https://doi.org/10.7554/eLife.45572.028
Figure 5—figure supplement 4—source data 1

CSA and sulfite are toxic to NSCLC cells.

https://doi.org/10.7554/eLife.45572.029
Figure 6
Download asset Open asset
Sulfitolysis is not required for the inhibition of proliferation by CDO1.

(a) Western blot analysis of GOT1 and β-ACTIN expression in parental (P) A549 and H460 cells, and GOT1 KO clones #1 and #2 for each cell line expressing empty pMXS (GOT1 -) and reconstituted with …

https://doi.org/10.7554/eLife.45572.031
Figure 6—source data 1

Sulfitolysis is not required for the inhibition of proliferation by CDO1.

https://doi.org/10.7554/eLife.45572.032
Figure 7 with 1 supplement
Download asset Open asset
CDO1-dependent cystine reduction limits NADPH availability for cellular processes.

(a) The NADPH/NADP + ratio was assayed following expression of CDO1Y157F (Y157F) or CDO1WT (WT) in A549 and H460 GOT1 KO cells expressing empty pMXS (GOT1 -) or reconstituted with pMXS-GOT1 (GOT1 …

https://doi.org/10.7554/eLife.45572.033
Figure 7—source data 1

CDO1-dependent cystine reduction limits NADPH availability for cellular processes.

https://doi.org/10.7554/eLife.45572.035
Figure 7—figure supplement 1
Download asset Open asset
Cystine uptake and reduction depletes NADPH.

(a) The NADPH/NADP+ ratio was assayed following growth of A549 cells in medium containing 200 µM or 0 µM cystine for 24 hr. The ratio was normalized to cells grown in 200 µM cystine. N = 3 …

https://doi.org/10.7554/eLife.45572.034
Figure 7—figure supplement 1—source data 1

Cystine uptake and reduction depletes NADPH.

https://doi.org/10.7554/eLife.45572.036
Figure 8
Download asset Open asset
Nrf2 activation promotes Cdo1 expression in murine lung tumors in vivo.

(a) Overall survival of Trp53flox/flox and KrasG12D; Trp53flox/flox mice expressing homozygous Keap1WT (WT/WT), heterozygous for Keap1R554Q (R554Q/WT) or homozygous for Keap1R554Q (R554Q/R554Q). (b) …

https://doi.org/10.7554/eLife.45572.037
Figure 8—source data 1

Nrf2 activation promotes Cdo1 expression in murine lung tumors.

https://doi.org/10.7554/eLife.45572.038
Figure 9
Download asset Open asset
Model: CDO1 antagonizes the growth and survival of KEAP1MUT cells by producing toxic products and depleting NADPH.

(Left) Elevated intracellular cysteine (CYS) stabilizes CDO1, leading to the production of CSA and SO32-. In turn, SO32- depletes cystine via sulfitolysis to produce CYS-SO3-, further limiting …

https://doi.org/10.7554/eLife.45572.039

Tables

Key resources table
Reagent type
(species) or
resource		DesignationSource or referenceIdentifiersAdditional information
Chemical
compound, drug		CycloheximideSigma Aldrich01810–1G
Chemical
compound, drug		PyridineSigma Aldrich270970–1L
Chemical
compound, drug		CuCl2Sigma Aldrich203149–10
Chemical
compound, drug		Methoxylamine
hydrochloride		Sigma Aldrich226904–5G
Chemical
compound, drug		Ethyl acetate
(LC-MS grade)		Sigma Aldrich34972–1 L-R
Chemical
compound, drug		Methanol
(HPLC grade)		Sigma Aldrich34860–1 L-R
Chemical
compound, drug		CysteineSigma AldrichC6852-25G
Chemical
compound, drug		CystineSigma AldrichC6727-25G
Chemical
compound, drug		L-Cysteine S-sulfateSigma AldrichC2196-25MG
Chemical
compound, drug		L-Cysteine
sulfinic acid		Sigma Aldrich270881–1G
Chemical
compound, drug		HypotaurineSigma AldrichH1384-100MG
Chemical
compound, drug		Na2SO3Sigma Aldrich71988–250G
Chemical
compound, drug		[34S]-Na2SO3,Sigma Aldrich753572
Chemical
compound, drug		DoxycyclineSigma AldrichD9891-1G
Chemical
compound, drug		[13C2,15N]-Glutathione
trifluoroacetate salt,		Sigma Aldrich683620
Chemical
compound, drug		GSSGSigma AldrichG4376-250MG
Chemical
compound, drug		Decitabine
(5-Aza-2’-deoxycytidine)		Sigma AldrichA3656-5MG
Chemical
compound, drug		Cumene
hydroperoxide		Invitrogencomponent
of C10445
Chemical
compound, drug		N-EthylmaleimideChem-Impex International00142
Chemical
compound, drug		[13C5]-glutamine,Cambridge Isotope LabsCLM-1822-H-0.25
Chemical
compound, drug		[13C3, 15N]-CysteineCambridge Isotope LabsCNLM-3871-H-0.25
Chemical
compound, drug		[13C3]-Cysteine,Cambridge Isotope LabsCLM-4320-H-0.1
Chemical
compound, drug		[D4]-Cystine,Cambridge Isotope LabsDLM-1000–1
Chemical
compound, drug		[13C2]-TaurineCambridge Isotope LabsCLM-6622
Chemical
compound, drug		[D5]-GlutathioneSanta Cruz Biotechnologysc-489493
Chemical
compound, drug		[D4]-HypotaurineCDN IsotopesH1384-100MG
Chemical
compound, drug		Formic acidFisher ChemicalMFX04405
Chemical
compound, drug		NaOHFisher ChemicalSS256500
Chemical
compound, drug		MSTFA + 1% TMCS
solution		Fisher ChemicalTS-48915
Chemical
compound, drug		HPLC-grade
water		Fisher ChemicalW5-1
Chemical
compound, drug		Acetonitrile
(HPLC grade)		Honeywell34967
Chemical
compound, drug		ErastinCayman Chemical17754
Chemical
compound, drug		β-lapachoneDr. David Boothman
Chemical
compound, drug		puromycinInvivogenant-pr-1
Chemical
compound, drug		blasticidinInvivogenant-bl-1
Strain, strain
background (mouse)		LSL-KrasG12D(Jackson et al., 2001)
Strain, strain
background (mouse)		Trp53flox(Marino et al., 2000)
Strain, strain
background (mouse)		Keap1R554Qthis paperA minigene containing
a cDNA encoding wild
-type exons 3–5, followed
by a SV40 polyA signal,
was inserted upstream
of endogenous exon 3
of the Keap1 gene. Codon
554 in endogenous
exon four was mutated
from arginine to glutamine
Recombinant
DNA reagent		Keap1R554Q
genotyping primers		this paperCommon (R) 5’-GCCACC
CTATTCACAGACCA-3’
Mutant (F) 5’-ATGGCCA
CACTTTTCTGGAC 3’
WT (F) 5’-GGGGGTAGA
GGGAGGAGAAT-3’		WT PCR product = 326 bp
Mutant PCR product
= 584 bp
Recombinant
DNA reagent		adenoviral-CreUniversity of IowaVVC-U of Iowa-5
Recombinant
DNA reagent		pRRL-CDO1this paperThe LT3GEPIR vector
backbone (Fellmann et al., 2013)
was obtained from
Johannes Zuber and the
MiR-E cassette was excised
to generate pRRL-GFP. GFP
was excised and replaced
with CDO1 using pQTEV-CDO1
(addgene# 31292) as a PCR
template.
Recombinant
DNA reagent		pRRL-CDO1 Y157Fthis paperThe enzyme inactive
mutant (CDO1Y157F) was
generated from pRRL-CDO1
by site-directed mutagenesis
of the wild-type protein.
Recombinant
DNA reagent		pQTEV-CDO1Addgene31292
Recombinant
DNA reagent		lentiCRISPR-V2(Shalem et al., 2014)
Recombinant
DNA reagent		lentiCRISPR-V2
mCdo1 #2		this paperprogenitor: lentiCRISPR-V2;
oligonucleotides for sgRNAs
targeting mCDO1 (#2F 5’-
caccgCGAGAGCAATCCCGCCGAGT-
3’, #2R 5’-aaacACTCGGCG
GGATTGCTCTCGc-3’)
Recombinant
DNA reagent		lentiCRISPR-V2
mCdo1 #3		this paperprogenitor: lentiCRISPR-V2;
oligonucleotides for sgRNAs
targeting mCDO1 (#3F
5’-caccgCGAAGAGCTCATGTAA
GATG-3’, #3R 5’-aaacCATCTT
ACATGAGCTCTTCGc-3’)
Recombinant
DNA reagent		lentiCRISPR-V2
hCDO1 #1		this paperprogenitor: lentiCRISPR-V2;
oligonucleotides for sgRNAs
targeting hCDO1 (F - 5’-caccgGAT
GCGGATCAGATCAGCCA-3’,
R - 5’-aaacTGGCTGATCT
GATCCGCATCc-3’)
Recombinant
DNA reagent		lentiCRISPR-V2
hCDO1 #2		this paperprogenitor: lentiCRISPR-V2;
oligonucleotides for
sgRNAs targeting hCDO1
(R - 5’-caccgCGAGAGCGACCCC
ACCGAGT-3’,R - 5’-aaacACTCG
GTGGGGTCGCTCTCGc-3’)
Recombinant
DNA reagent		pLX317-NRF2Dr. Alice Berger,
(Berger et al., 2016)
Recombinant
DNA reagent		pLX317-NRF2T80KDr. Alice Berger,
(Berger et al., 2016)
Recombinant
DNA reagent		pLX317-emptythis paperpLX317-empty was
generated from pLX317
-NRF2 by site
directed mutagenesis.
Recombinant
DNA reagent		pLenti KEAP1WTthis paperThe KEAP1WT cDNA was
provided by Dr. Christian
Metallo (Zhao et al., 2018);
pLenti-GFP-blast was
generated from pLenti-
GFP-puro (addgene #17448)
Recombinant
DNA reagent		pLenti KEAP1C151Sthis paperThe KEAP1C151S cDNA
was provided by Dr. Christian
Metallo (Zhao et al., 2018);
pLenti-GFP-blast was
generated from pLenti-GFP
-puro (addgene #17448)
Recombinant
DNA reagent		pLenti KEAP1C273Sthis paperThe KEAP1C273S cDNA
was provided by Dr. Christian
Metallo (Zhao et al., 2018);
pLenti-GFP-blast was
generated from
pLenti-GFP-puro
(addgene #17448)
Recombinant
DNA reagent		plentiCRISPR-sgGOT1addgene72874
Recombinant
DNA reagent		pMXS-GOT1addgene72872
Recombinant
DNA reagent		pMXS-emptythis paperpMXS-empty was generated
from pMXS-GOT1 by
site-directed mutagenesis.
Recombinant
DNA reagent		pCMV-dR8.2 dvpraddgene8455
Recombinant
DNA reagent		pCMV-VSV-Gaddgene8454
Cell line
(Homo-sapiens)		Lenti-X 293TTakara632180
Cell line
(Homo-sapiens)		Phoenix-AMPHOATCCCRL-3213RRID:CVCL_H716
Cell line
(Homo-sapiens)		Calu3Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_0609
Cell line
(Homo-sapiens)		H1581Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1479
Cell line
(Homo-sapiens)		H1975Dr John Minna,
Hamon Cancer Center
Collection (University of
Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1511
Cell line
(Homo-sapiens)		H2087Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1524
Cell line
(Homo-sapiens)		H2347Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1550
Cell line
(Homo-sapiens)		H1792Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1495
Cell line
(Homo-sapiens)		H1944Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1508
Cell line
(Homo-sapiens)		H322Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_1556
Cell line
(Homo-sapiens)		H460Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_0459
Cell line
(Homo-sapiens)		HCC15Dr John Minna,
Hamon Cancer Center
Collection (University
of Texas-Southwestern
Medical Center)		(DeNicola et al., 2015)RRID:CVCL_2057
Cell line
(Homo-sapiens)		H2009ATCCCRL-5911RRID:CVCL_1514
Cell line
(Homo-sapiens)		H1299ATCCCRL-5803RRID:CVCL_0060
Cell line
(Homo-sapiens)		H1993ATCCCRL-5909RRID:CVCL_1512
Cell line
(Homo-sapiens)		H441ATCCHTB-174RRID:CVCL_1561
Cell line
(Homo-sapiens)		A549ATCCCCL-185RRID:CVCL_0023
Cell line
(Homo-sapiens)		NRF2 KO A549Dr. Laureano
de la Vega		(Torrente et al., 2017)
Cell line (Mus
musculus)		mouse embryonic
fibroblasts		this paperMEFs were isolated
from E13.5–14.5 day
old embryos
Commercial
assay or kit		JetPRIMEVWR89129–922
Commercial
assay or kit		ImmPRESS HRP
anti-rabbit kit		Vector LabsMP-7451
Commercial
assay or kit		E.Z.N.A. Total RNA Kit IOmega BiotekR6834-02
Commercial
assay or kit		PrimeScript
RT Master Mix		TakaraRR036A
Commercial
assay or kit		Taqman gene
expression assays
- mCdo1		Thermo Fisher4448892,
Mm00473573_m1
Commercial
assay or kit		Taqman gene
expression assays
- hCDO1		Thermo Fisher4448892,
Hs01039954_m1
Commercial
assay or kit		Taqman gene
expression assays
- mActb		Thermo Fisher4448892,
Mm02619580_g1
Commercial
assay or kit		Taqman gene
expression assays
- hACTB		Thermo Fisher#4333762F
Commercial
assay or kit		CellTiter-GloPromegaG7571
Commercial
assay or kit		DC protein assayBiorad500112
AntibodyNRF2 (Rabbit mAb)Cell Signaling
Technologies		12721RRID:AB_2715528;
1:1000 WB
AntibodyHSP90 (Rabbit pAb)Cell Signaling
Technologies		4874 sRRID:AB_2121214;
1:5000 WB
AntibodyTXN1 (Rabbit mAb)Cell Signaling
Technologies		2429SRRID:AB_2272594;
1:1000 WB
Antibodyα-tubulin (Mouse mAb)Santa Cruzsc-8035, clone TU-02RRID:AB_628408;
1:500 WB
Antibodyβ-actin (Mouse mAb)Thermo FisherAM4302, clone AC-15RRID:AB_2536382;
1:100,000 WB
AntibodyCDO1 (Rabbit pAb)Abcamab53436RRID:AB_940958;
1:1000 WB, discontinued,
verified with Sigma
CDO1 antibody
AntibodyxCT (Rabbit pAb)Abcamab37185RRID:AB_778944;
1:1000 WB
AntibodyGOT1 (Rabbit pAb)BiovisionA1272RRID:AB_2801348;
1:1000 WB
AntibodyCSAD (Rabbit pAb)LSBioC375526RRID:AB_2801349;
1:1000 WB
AntibodyAnti-NQO1
antibody
(Rabbit pAb)		Sigma AldrichHPA007308RRID:AB_1079501;
1:100, IHC; 1:1000 WB
AntibodyAnti-CDO1
antibody
(Rabbit pAb)		Sigma AldrichHPA057503RRID:AB_2683451;
1:100, IHC
Otherdialyzed FBSSigma AldrichF0392
OtherCysteine/cystine,
methionine and
glutamine free RPMI		MP Biomedicals91646454
OtherCysteine/cystine,
methionine, pyruvate
and glutamine free
DMEM		Gibco21013024
Software,
algorithm		MZmine 2(Pluskal et al., 2010)Version 2.30
Software,
algorithm		Thermo Xcaliber
Qual Browser		Thermo FisherVersion 4.0.27.19
Software,
algorithm		EI Mavenhttps://elucidatainc.github.io/ElMavenVersion 0.3.1
Software,
algorithm		Agilent Mass Hunter
Workstation Software
- Qualitative Analysis		AgilentVersion B.07.00
Software,
algorithm		Image Scope softwareAperio

Additional files

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https://doi.org/10.7554/eLife.45572.040

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