A B 12 *** *** *** *** *** 12
10 10 r=0.62
expression (log2) expression p<0.0001 8 n=1063 MEN1 MEN1 expression (log2) expression MEN1 8 -2 0 2 4 MEN1 Copy Number ((log2)-1)
like (n = 8) - -like (n = 89) Normal (n = 61) enriched (n = 54) Basal - Normal Luminal A (nLuminal = 221) B (n = 122) HER2 C D
(%) (%) (%) (%) ERa+ ERa- 100 ERa+ ERa- 100 100 100 75 75 p<0.01 75 p=n.s. p<0.0001 75 p=n.s.
50 50 50 50
CN>2 (n=81) CN>2 (n=21) 25 High (n=207) 25 High (n=74) 25 25 CN=2 (n=830) CN=2 (n=309) Low (n=207) Low (n=74) 0 0 CN<2 (n=162) 0 CN<2 (n=49) Disease free probabilityDisease free 0 0 100 200 300 0 100 200 300 probabilityDisease free 0 100 200 300 0 100 200 300 Months Months Months Months
Figure S1. Increased MEN1 expression and copy number are related to poor prognosis of ERa+ breast cancer patients. (A) MEN1 expression in different breast cancer PAM50 subtypes in TCGA breast cancer cohort. Significant levels were based on Duncan’s multiple range tests (***P < 0.001). (B) Scatter plot and linear regression of MEN1 expression and copy number in TCGA breast cancer cohort. Significant levels were based on Pearson correlation coefficient. (C) and (D) Kaplan-Meier survival analyses and significant levels were based on logrank tests from Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) cohort based on MEN1 expression (c) or copy number (D). A rtTA Doxycycline BirA Menin Biotin MAPs
B
T47D EIF2B5 KMT2D
CBX4
AFF4 NOC2L GATA3 NOLC1 EP400 LIG3
GTF TLE3 ECPAS 3C1 WAPL KDM3B
SF1 MED12 NUP153 MKI67 SART1 Menin ZFP36 SNRPD3 L2 SMC HD1 MSH2 KMT2A SRSF5 EMSY POLR BPTF 2A
PURB STAG1 TP53 I11 LENG8 ZC3H18 FOXK1
KPNA6
FRG1 C Glycolytic enzyme
7 ALDOA 7 TPI1 3 ENO2
Menin 0 0 0
7 7 3
KMT2A 0 0 0 chr16 ⎮30071000 ⎮30076000 ⎮30081000 ⎮ chr12 ⎮6971000 ⎮6975000 ⎮6979000 chr12 ⎮7015000 ⎮7024000 ⎮7033000 ⎮
D OXPHOS complex I
NDUFA11 NDUFB5 7 7 7 NDUFB6
Menin 0 0 0
7 7 7
KMT2A 0 0 0 chr19 ⎮5885000 ⎮5894000 ⎮5903000 ⎮5912000 chr3 ⎮179319000 ⎮179325000 ⎮179331000 ⎮ chr9 ⎮32561000 ⎮32567000 ⎮325730000 ⎮
NDUFS6 NDUFS7 7 7
Menin 0 0
7 7
KMT2A 0 0 chr5 ⎮1799000 ⎮1803000 ⎮1807000 chr19 ⎮1381000 ⎮1385000 ⎮13889000
OXPHOS complex II OXPHOS complex III OXPHOS complex IV SDHA UQCRB COX6B1 7 7 7
Menin 0 0 0
7 7 7
KMT2A 0 0 0 chr5 ⎮214000 ⎮220000 ⎮226000 ⎮ chr8 ⎮97240000 ⎮97245000 ⎮97250000 ⎮ chr19 ⎮36137000 ⎮36141000 ⎮36145000
OXPHOS complex V COX14 ATP5F1D ATP5PB 7 14 7
Menin 0 0 0
7 14 7
KMT2A 0 0 0 chr12 ⎮50502000 ⎮50507000 ⎮50512000 ⎮ chr19 ⎮1240500 ⎮1242100 ⎮1243700 ⎮124530 chr1 ⎮111984000 ⎮111991000 ⎮11199000 ⎮
Figure S2. The BioID system, network analysis of 35 MAPs in T47D and in silico analysis of anti-menin and anti-KMT2A ChIP-seq. (A) Schematic illustration of biotin labeling of MAPs using BioID proximity-dependent biotinylation with a doxycycline-inducible biotin ligase BirA-MEN1 fused gene. (B) Network analysis of 35 MAPs in T47D cells. The distance between menin and MAPs represented the protein quantitative ratio of each MAP and menin. MAPs marked in blue were further validated by WES. (C) In silico analysis of anti-menin and anti-KMT2A ChIP-seq data indicates that anti-Menin immunoprecipitation enriched the promoter and gene body DNA sequence of the representative glycolytic genes and (D) OXPHOS genes in MCF-7 cells. C A GATA median values tumors breast OXPHOS Heatmaps Expression Figure MEN1 expression OXPHOS (n=87) Tumor OXPHOS (n=87) Normal 10 11 12 13
8 9 MEN1 CBX4 8 3 ZC3H18 . Group 3: Low 3: Group Low 4: Group cancer ) S
of EIF2B5 . The
expression SNRPD3 3 SART1 . genes both 9
of NOC2L correlation
KMT2A FRG1 tumors ECPAS gene - - cohort
Low High MSH2 10 MKI67 genes’
expression NOLC1 (
A SF1 ZFP36L2 ) are 11 expression SRSF5 of .
or LENG8 ( Group 1: High 1: Group Group 2: High 2: Group C
relationship GATA3 MEN grouped
expression TLE3 glycolytic ) 12 LIG3
Scatter GTF3C1 TP53I11
1 FOXK1 -
- KPNA6 Low High PURB and
correlations MED12 into POLR2A plot
genes NUP153 . of SMCHD1 the
Similar STAG1 4 MAPs KMT2A of
groups EMSY BPTF other MEN
( WAPL B AFF4
) KDM3B and of groupings KMT2D in 1 EP400 ( three 1 37 - and normal 0.7 B - corr. coef. corr. OXPHOS/glycolytic 4 0 ) MAPs 0.7 as Tumor Normal
KMT Glycolysis (n=77) Glycolysis (n=77) MAP SNRPD3
in MEN1 are and ZC3H18 2
Figure NOC2L (including A SART1
genes FRG1 also tumor expression EIF2B5 SRSF5 LENG8
3 GATA3 applied
E TLE3
( CBX4 samples MED
according GTF3C1 genes MEN TP53I11 FOXK1 SF1 in 12 ZFP36L2
1 NOLC1 based . TCGA , MKI67 )
( MSH2 from WAPL A genes NUP153 to
) SMCHD1 STAG1 and
median MED12 on TCGA breast KPNA6 PURB with and ( KMT2A the B WAPL
) BPTF
, EMSY AFF4 KDM3B KMT2D EP400 LIG3 POLR2A ECPAS A T47D MCF-7 T47D MCF-7 T47D MCF-7 T47D MCF-7
WAPL WAPL KMT2A KMT2A MED12 GATA3 GATA3 MED12 sh sh sh Vehicle sh Vehicle Vehiclesh Vehiclesh Vehicle Vehicle Vehiclesh Vehiclesh KMT2A MED12 WAPL GATA3-FL GATA3-SP b-Actin b-Actin b-Actin b-Actin
B T47D MCF-7 C T47D MCF-7
KMT2A GATA3 KMT2A GATA3 KMT2A KMT2A GATA3 Vehiclesh Vehiclesh Vehiclesh Vehiclesh Vehiclesh VehicleshGATA3 Vehiclesh Vehiclesh NDUFA13 NDUFA13 ALDOA ALDOA NDUFS7 NDUFS7 ALDOC ALDOC NDUFS8 NDUFS8 ENO1 ENO1 NDUFB7 NDUFB7 PFKL PFKL NDUFV1 NDUFV1 PFKP PFKP SDHA SDHA PGK1 PGK1 SDHB SDHB TPI1 TPI1 SDHC SDHC KMT2A KMT2A 0 1 2 SDHD SDHD GATA3 GATA3 Relative expression
D Oligo FCCP Rot+AA E 20 T47D Vehicle T47D Vehicle T47D shMED12 T47D Vehicle 20 15 15 1515 T47D shMEN1 T47D shKMT2A T47D shWAPL T47D shGATA3
1515 Spare 10 Capacity 1010 10 1010 /min/1000 cells) /min/1000 cells) ATP Maximal 55 5 5 55 Basal mpH Proton Leak mpH Non-Mitochondrial Respiration 0 00 00 0 OCR ( 00 2020 40 60 80 OCR ( 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 Time (min) Time (min) Time (min) Time (min)
F MCF-7 Vehicle MCF-7 Vehicle MCF-7 shMED12 MCF-7 Vehicle MCF-7 Vehicle MCF-7 shMEN1 MCF-7 shKMT2A MCF-7 shWAPL MCF-7 shGATA3 15 15 15 20
15 10 10 10 10 /min/1000 cells) 5 5 5 5 mpH
0 0 0 0 OCR ( 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 G Time (min) H Time (min) Time (min) Time (min) 4 4 Glucose Oligo 2-DG 4 T47D Vehicle T47D Vehicle T47D shMED12 4 T47D Vehicle T47D shMEN1 T47D shKMT2A T47D shWAPL T47D shGATA3 3 3 3 3 Glycolytic 2 Reserve 2 2 2
Glycolysis /min/1000 cells) /min/1000 cells) Glycolytic 1 1 1 Capacity mpH 1 mpH Non-Glycolytic Acidification 0 0 0 0
0 20 40 60 80 ECAR ( 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 ECAR ( I Time (min) Time (min) Time (min) Time (min) MCF-7 Vehicle MCF-7 Vehicle MCF-7 shMED12 MCF-7 Vehicle MCF-7 Vehicle 4 4 MCF-7 shMEN1 4 MCF-7 shKMT2A MCF-7 shWAPL 4 MCF-7 shGATA3
3 3 3 3
2 2 2 2 /min/1000 cells)
1
mpH 1 1 1
0 0 0 0
ECAR ( 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 Time (min) Time (min) Time (min) Time (min) Figure S4. Gene and protein expressions and extracellular-flux analysis in MAP shRNA knockdown cells. (A) WES of total lysate from T47D or MCF-7 cells subject to vehicle or MAP shRNA lentivirus infection. (B) and (C) Heat maps of RT-qPCR of OXPHOS genes (B) or glycolytic genes (C) in T47D or MCF-7 cells. The expression in vehicle control is normalized as 1 (n = 3). (D) Seahorse mitochondrial stress test profile. (E) and (F) Line charts of OCR for mitochondrial functions in T47D (E) and MCF-7 (F) cells. (G) Seahorse glycolytic stress test profile. (H) and (I) Line charts of ECAR for glycolytic functions in T47D (H) and MCF-7 (I) cells. Data are presented as mean±S.D. (n = 15-20 technical-replicate wells). A C T47D MCF-7 D T47D MCF-7 1.21.2 T47D MCF-7 *** 0.80.8 503 0 h503 1 h503 3 h503 6 h -503 0 h503 1 h503 3 h503 6 h503 72 h 503 0 h503 1 h503 3 h503 6 h503 72 h 503 0 h503 1 h503 3 h503 6 h503 72 h ** - - - - -503 72 h ------** MI MI MI MI MI MI MI MI MI MI MI MI MI MI MI MI MI MI MI MI 0.40.4 NDUFS7 ALDOA NDUFA11 Cell viability (%) viability Cell ALDOC 0.00.0 SHDA ENO1 0 0.5 1 1.5 2 2.5 SDHB PFKL MI-503 (µM) CYC1 ctrl PFKP B MI(1) MI(2) COX6C PGK1 T47D UQCR11 TPI1 ATP5D 0 1 2 Relative expression Input IP:Menin IP:IgG WAPL
E T47D MI-503 0h 15MCF-7 MI-503 0 h MCF-7 MI-503 3 h MCF-7 DMSO 72 h T47D MI-503 1h MCF-7 MI-503 1 h MCF-7 MI-503 6 h MCF-7 MI-503 72 h T47D MI-503 3 h T47D DMSO 72 h Oligo FCCP Rot+AA T47D MI-503 6 h T47D MI-503 72 h Oligo FCCP Rot+AA
10 10 10 10 Oligo FCCP Rot+AA Oligo FCCP Rot+AA
5 5 5 5 /min/1000 cells) mpH
OCR ( 0 0 0 0 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 Time (min) Time (min) Time (min) Time (min) F T47D MI-503 0 h T47D DMSO 72 h MCF-7 MI-503 0 h MCF-7 MI-503 3 h MCF-7 DMSO 72 h T47D MI-503 1h T47D MI-503 72 h MCF-7 MI-503 1 h MCF-7 MI-503 6 h MCF-7 MI-503 72 h T47D MI-503 3 h Glucose Oligo 2-DG Glucose Oligo 2-DG T47D MI-503 6 h 4 4 4 4
Glucose Oligo 2-DG Glucose Oligo 2-DG 3 3 3 3
2 2 2 2 /min/1000 cells)
mpH 1 1 1 1
0 0
ECAR ( 0 0 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 Time (min) Time (min) Time (min) Time (min)
Figure S5. Gene expressions and extracellular-flux analysis in cells treated with MI-503. (A) Cell viability of T47D and MCF-7 cells under different dosages of MI-503 for 3 days (n= 5). (B) Co-immunoprecipitation of WAPL, another MAP, with menin pulled down by anti-menin but not anti-IgG. (C) and (D) Heat maps of relative expression of OXPHOS (C) and glycolytic (D) genes by RT-qPCR in T47D or MCF-7 cells subject to 1 µM MI-503 for 0, 1, 3, 6, 72 h, or DMSO for 72 h. Gene expressions post MI-503 treatment for 1, 3, and 6 h were normalized to that of the 0 h control; gene expressions post MI-503 treatment for 72 h were normalized to that of DMSO treatment for 72 h. The expressions in 0 h of MI-503 treated cells and 72 h of DMSO treated cells were normalized as 1 (n = 3). (E) and (F) Line charts of extracellular-flux analysis for mitochondrial (E) and glycolytic (F) functions in T47D or MCF-7 cells treated by MI-503 for 0, 1, 3, 6, 72 h, or DMSO for 72 h. Data are presented as mean ± S.D. (n = 15-20 technical-replicate wells). A CTCs isolation C Circulation system
PBMCs Immunofluorescent CD45 staining (7 days) depletion
Single-cell RT-qPCR Blood Ficoll Ultra-low sample isolation adherent plate
B DAPI CD44 Pan-CK ALDH1 Merge P8 Cell viability assay
Circulation
Seahorse assays P9
DAPI NANOG OCT4 Merge
P8 D 1.21.2 T47D MCF-7
P9 0.80.8 *** *** *** *** DAPI EpCAM CD45 Merge 0.40.4 Cell viability Cell
P8 0.00.0 0 1 2 0 1 2 1 2 3 4 5 6 7 8 Circulation (h) P9
Figure S6. CTCs isolation and in vitro circulation system. (A) Schematic workflow of CTC isolation, expansion, and subsequent experiments. (B) Representative images of CTCs staining with stem cell or epithelial markers. Scale bar, 10 µm. (C) Scheme of the circulation system and experimental assays. (D) Bar chart of cell viability of T47D and MCF-7 cells after circulation (n = 5 technical-replicate wells). Data are presented as mean ± S.D. Unpaired two-tailed Student’s t-test was used for statistical significance determination of 0 h and each treatment (***P < 0.001). MCF7
1.0 P < 0.0001
****
0.5
Relative ATP production rate 0.0 Vehicle ShMEN1
Figure S7. Validation of ATP quantitation by Seahorse Seahorse XFe96 analyzer using CellTiter-Glo Luminescent Cell Viability Assay Kit. ATP reduction is observed in MCF7 cells treated with menin shRNA, consistent with the measurement result by Seahorse in Fig. 4A. Fig. 1b Fig. 2f, BirA-Menin Fig. 2f, KMT2A Fig. 2f, MED12 T47D MCF-7 Fig. 2A Fig. 2B T47D MCF-7 T47D MCF-7 T47D MCF-7
Fig. 2f, WAPL Fig. 2f, GATA3 Fig. 2f, LaminA/C; GAPDH Fig. 5a, Menin Fig. 5a, KMT2A T47D MCF-7 T47D MCF-7 T47D MCF-7 T47D MCF-7 T47D MCF-7
Fig. 5a, GATA3 Fig. 5a, LaminA/C Fig. 5b, Menin Fig. 5b, KMT2A; GATA3 T47D MCF-7 T47D MCF-7 T47D MCF-7 Fig. 5b, KMT2A (T47D) Fig. 5b, GATA3 (T47D) (MCF-7)
Fig. S4a, KMT2A b-Actin Fig. S4a, MED12 b-Actin Fig. S4a, WAPL b-Actin T47D MCF-7 T47D MCF-7 T47D MCF-7 T47D MCF-7 T47D MCF-7 T47D MCF-7
Fig. S4a, GATA3 b-Actin T47D MCF-7 T47D MCF-7
Figure S8. The original WES images. Table S1. Biotin labelled nuclear proteins in T47D and MCF-7 cells. Related to Figure 2C. T47D MCF-7 ACIN1, AFF4, BPTF, BTAF1, CCNT1, CHD8, Class 1: DHRS7B, EIF2B5, EIF3C, ELMSAN1, GTF2F2, AFF4, BPTF, CHD1, CHD3, EIF2B5, Basal GTF3C1, HP1BP3, INTS1, INTS9, LIG3, MED1, EIF5B, GRHL2, GTF3C1, HCFC1, transcription MED12, MED14, NELFE, NOC3L, ORC2, HIRA, INTS8, LIG3, MED12, factor PAF1, PAPOLA, POLA1, POLR2A, POLR2B, MED24, NCOR1, POLR1D, complexes RFC5, SENP3, SUPT16H, TAF7, TOX4, WAC, POLR2A, POLR2E, SMC6, WDHD1. WDR82, ZC3H4, ZHX3, ZMYND8. Class 2: ARID1B, BAZ1B, DCAF13, DDB1, EMSY, ARID1A, EMSY, FANCI, MDC1, MSH2, NBN, PMS2, RBBP8, RIF1, SSBP1, DNA repair MSH2, TNKS1BP1. complexes XRCC1. C17orf85, C19orf43, CMTR1, CPSF6, DDX10, DDX23, DDX27, DDX41,DDX42, DDX46, ATXN2L, CNOT2, CPSF2, DCP1A, DDX54, DDX56, DHX36, DHX38, DHX40, EDC4, ESRP1, FRG1, GEMIN4, Class 3: DIS3, EIF2AK2, FIP1L1, FRG1, FUBP1, GEMIN6, KHDRBS1, KHSRP, MCM3AP, MRTO4, NHP2L1, NUFIP2, PARN, mRNA PCID2, RANGAP1, SART1, SF1, PLRG1, PRCC, PRPF31, PRPF6, RAE1, modeling; RNA SF3A2, SF3B2, SKIV2L2, SNRPD3, RBM10, RBM17, RBM26, RBM28, RBM7, helicase; SRRM2, SRSF3, SRSF5, THOC1, SART1, SART3, SF1, SF3A1, SF3B5, SF3B6, spliceosomes THRAP3, TRA2A, UPF2, ZCCHC8, SMN1, SNRPD1, SNRPD3, SRBD1, SRSF5, ZFP36L2. TFIP11, TTC37, WDR33, XAB2, ZFP36L2, ZNF638. Class 4: DRG1, DRG2, ECM29, KPNA6, MISP, MKI67, ECM29, EHD4, KPNA6, MAD1L1, NDC1, NSFL1C, NUP107, NUP153, NUP188, MKI67, MYH10, NOP14, NUP153, Nuclear scaffold NUP205, NUP85, PPP1R10, SMC1A, SMC4, NUP210, NUP88, PLK1, POM121C, complexes SMCHD1, SUN2, SURF6, TUBB6, TUBGCP2, SAFB2, SEC13, SMCHD1, SYMPK, relate WDR3, WDR46. TPX2. Class 5: CHAMP1, NCAPD2, NCAPD3, PDS5A, Site-specific STAG1, WAPL. transcription PDS5B, POGZ, STAG1, TOP1, TOP2B, WAPL. factors ADNP, AQR, ARFGEF1, BDH1, CBX4, CRK, CBX4, CHCHD3, DIDO1, ERBB2IP, Class 6: DNAJA3, FOXK1, FOXK2, GATA3, GREB1, FOXK1, G3BP1, GATA3, GATAD2B, JAK1, MKL2, MLLT11, MYBBP1A, NCOA6, GSE1, ILF3, MBD3, MYEF2, Cohesin related NFIC, NKRF, PARP14, PEG10, PURB, RFX1, PPM1D, PURB, SAMD1, SPEN, complexes and SBNO1, SCML2, SCYL1, SENP1, SIN3A, TCF25, TLE3, TP53BP1, TP53I11, mediators STAT2, TCF20, TLE3, TP53I11, YAP1, YY1AP1, TRPS1, TSC2, ZC3H18, ZMYM2, ZC3H18, ZNF148, ZNF281, ZNF687. ZNF384, ZNF644. BAZ2A, EP400, HIST1H2BB, Class 7: EP400, EZH2, GRWD1, HDAC2, JMJD1C, HIST1H2BC, KAT6B, KDM3B, Histone KDM2A, KDM3B, KMT2A, KMT2D, NOC2L, KMT2A, KMT2D, NOC2L, OGT, modifying PHF8, RSBN1, RSBN1L, SETD1A, PAXIP1, SAP130, SMARCAL1, enzymes SMARCAD1, TRRAP, WIZ. SMARCC2, SRCAP, STK19, SUDS3. ACOT13, BCLAF1, CRNKL1, CUL4B, CUL5, DARS2, DERA, EARS2, EBP, FAM120C, FBXL6, FDPS, GNL2, GPATCH1, IMP4, IPO11, Class 8: ITPR2, KARS, KIF13B, L1RE1, LENG8, NFS1, NOL6, NOLC1, NOP2, PCNP, PDCD6, AP3D1, BAG2, BOLA2, CBS, DUT, tRNA PPP2R5D, PPT1, PRKACG, PRKCD, PSPC1, IMPDH2, LAS1L, LENG8, NOLC1, modification, QIL1, RANBP3, RBBP6, RNF213, RPF2, PARL, PISD, PUS1, SRRT, UNC50. ubiquitination, RPP30, RPP38, RSL1D1, SRP14, SUGT1, and others TAOK2, TCOF1, TRMT1L, TRMT5, TRNT1, UBE2L6, UBE3A, UBE4B, UBR4, UBXN7, UGDH, UNC45A, UTP6, WDR75, YARS2, YIF1B, YLPM1. Table S2. shRNA sequences used in this study. shMEN1 CCGGGCTGCGATTCTACGACGGCATCTCGAGATGCCGTCGTAGAATCGCAG CTTTTTG shKMT2A CCGGCGCCTAAAGCAGCTCTCATTTCTCGAGAAATGAGAGCTGCTTTAGGC GTTTTT shMED12 CCGGGCAGAGAAATTACGTTGTAATCTCGAGATTACAACGTAATTTCTCTGCT TTTT shWAPL CCGGACGTACTGTAGGGCCAATAAACTCGAGTTTATTGGCCCTACAGTACGT TTTTTG shGATA3 CCGGCATCCAGACCAGAAACCGAAACTCGAGTTTCGGTTTCTGGTCTGGAT GTTTTT Table S3. RT-qPCR and Bio-mark primers used in this study. Gene F1 (5’- sequence -3’) R1 (5’- sequence -3’) R2 (5’- sequence -3’) MEN1 CCAGACAGTCAATGCCG CAGGTCAATGGAAGGGTT TGAACGCCACCTCCATC AAATGAGAGTAATGATAGG KMT2A GTGGAGGTGGAGACGAG TGGGGGGCACTTTGTAGA AGAAGC MED12 TCAGTCAACAAGCACGC AGGAGGATGGTCTGTAGG GGGCACATAAGCAGGTC GGGTGGGTAGAAGGAAG TCAAAAATAGGAAGACAAT WAPL GTGACAAATCGGGGCTC AAA AGAAGGC GGATTTGCTAGACATTTTT TGCCTTCCTTCTTCATAGT GATA3 CCACCACAACCACACTC CGG CAG CAATCTTCAGCACACAAC ALDOA TCAAATCCAAGGGCGGT CTTGGGTGGTAGTCTCG G GCCATCCTGTTCTGACTT ALDOC TTCACCTGGCACCAACT GCTCCCTATCCTCCCATC CTA CCAATAGCAGTCTTCAGC ENO1 GGAAAGATGCCACCAATG CCCAGACCTGAAGAACT AG PFKL CGACTGGCTGTTCATCC GAGATGGGCTTCCCGTT CCCACGGCTCCGAGTCT PFKP GGAGAAGCACGAGGAGT GCAGGTGTCGGTGATAGT AGTGTTCAGGGCGGTGT PGK1 TCGGGCTAAGCAGATTGT CAAGTGGCAGTGTCTCC GCTTTCACCACCTCATCC GAGCAGACAAAGGTCATC CAGTCTTGCCAGTACCAA TPI1 TGGACTTCAGCCATCCTC G T AAAACTAGAAGTGATACCT NDUFA7 ACTACTTTGCCTCCTTGG GCCTGGGAAACATGGTG GAGC AGTGAAACCCTGTCTCTAC NDUFA11 GTGACATGATCTCGGCTC TTCTCATGCCTCAGCCTC TAT GTACAGCTCCCCGATCAA NDUFA13 TACAGGCAGAAACCGACC GAACACAGACTCCCCCAC G NDUFB7 CACACAGCAGGAGATGAT GCAGTAGTCCCAGTCGT CGCTTGCACTTGAGCAG NDUFS7 GCACACTCACCAACAAGA GATGTAGATGTCCACGGG CATGGAGACCACGTAGCG ATCCCGAGATGGACATGA NDUFS8 GTAGTTGATGGTGGCCG GAACAGGTAGCTCAGGGT A NDUFV1 CTGTGGAGGAGGAGATGT GGGGATCAGTGGGGTAG AGTGGGGTAGACGAGCC CATTCCCACCAACTACAAG SDHA GACAACCAGGTCCAAGAG TGTACCGAGGCACAGGC G GCATAGAAGTTGCTCAAA GTGTGGAAGAGGGTAGAT SDHB GAGAAGGCATCTGTGGC TCG TT ACATACAGTGGTTTGGGT CCAGAAATGACCAAGAGA SDHC GTGGGTAGGGTTGAGGGA G AAG CTTATCTGTTATTTCTTCC SDHD AACTATCACGATGTGGGC AAGCAGAGGCAAAGAGG TTATTGTGAG CYC1* CGGAGGTGGAGGTTCAA CACGATGTAGCTGAGGTC CGTATCAAGGACAGTAACT ATCAGCCACACGAAACTT COX6C* ACCA A CCCAGATACATCACTCCC UQCR11* GCTGACAGAAACTGCCTA AA CATGTACGTTGCTATCCAG CTCCTTAATGTCACGCAC CTCCTTAATGTCACGCAC ACTB GC GAT GAT
*RT-qPCR only, F1 and R1 was used. For ACTB, same primer set was used for outer and inner primer.