KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal anti-PGC1α antibody, WB, dil:1/1,000 | Abcam | Cat#ab54481; RRID: AB_881987 |
| Rabbit monoclonal anti-Phospho-AMPKα (Thr172) (40H9) antibody, WB, dil:1/1,000 | Cell Signaling Technology | Cat#2535; RRID: AB_331250 |
| Rabbit polyclonal anti- PPARα antibody, WB, dil:1/1,000 | Abcam | Cat#ab24509; RRID: AB_448110 |
| Rabbit monoclonal anti-AMPKα (D5A2) antibody, WB, dil:1/1,000 | Cell Signaling Technology | Cat#5831; RRID: AB_10622186 |
| Rabbit monoclonal anti-Acetyl-CoA Carboxylase (C83B10) antibody, WB, dil:1/1,000 | Cell Signaling Technology | Cat#3676; RRID: AB_2219397 |
| Rabbit monoclonal anti-Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) antibody, WB, dil:1/1,000 | Cell Signaling Technology | Cat#11818; RRID: AB_2687505 |
| Rabbit monoclonal anti-Total OXPHOS antibody, WB, dil:1/500 | Abcam | Cat#ab110413; RRID: AB_2629281 |
| Rabbit monoclonal anti-β-Actin (D6A8) antibody, WB, dil:1/3,000 | Cell Signaling Technology | Cat#8457; RRID: AB_10950489 |
| Rabbit monoclonal anti-GAPDH (D16H11) antibody, WB, dil:1/3,000 | Cell Signaling Technology | Cat#5174; RRID: AB_10622025 |
| Rabbit polyclonal anti-Hsp90 antibody, WB, dil:1/2,000 | Cell Signaling Technology | Cat#4874; RRID: AB_2121214 |
| Rabbit monoclonal anti-α-Actinin (D6F6) antibody, WB, dil:1/2,000 | Cell Signaling Technology | Cat# 6487; RRID: AB_11179206 |
| Rabbit monoclonal anti-β-Tubulin (9F3) antibody, WB, dil:1/2,000 | Cell Signaling Technology | Cat#2128; RRID: AB_823664 |
| Rabbit polyclonal anti-Ceruloplasmin antibody, WB, dil:1/1,000 | Thermo Fisher Scientific | Cat#PA5-14225; RRID: AB_2085718 |
| Mouse monoclonal anti-SCO1 (D-1) antibody, WB, dil:1/500; IP, dil:1/100 | Santa Cruz Biotechnology | Cat#sc-398001; |
| Rabbit polyclonal anti-SCO1 antibody, WB, dil:1/1,000; IF, dil:1/200 | Proteintech | Cat#12614-1-AP; RRID: AB_2184641 |
| Mouse anti-β-Actin antibody, IF, dil:1/200 | Earthox | Cat#E021090-02; |
| Alexa Fluor 488 Secondary Antibody, IF, dil:1/200 | Thermo Fisher Scientific | Cat#A32731; RRID: AB_2633280 |
| CoraLite®488-Conjugated AMPK Alpha Monoclonal Antibody | Proteintech | Cat# CL488-66536; RRID: AB_2883352 |
| Rabbit monoclonal anti-LKB1(D60C5) antibody, WB, dil:1/1,000 | Cell Signaling Technology | Cat#3047; RRID: AB_2198327 |
| Rabbit polyclonal anti-acetylated-Lysine antibody, WB, dil: 1/1,000 | Cell Signaling Technology | Cat#9441; RRID: AB_331805 |
| Mouse monoclonal anti-Myc-Tag antibody, WB, dil:1/1,000; IP, dil: 1/250 | OriGene | Cat#TA150121; RRID: AB_2622266 |
| Mouse monoclonal anti-DYKDDDDK tag antibody, WB, dil:1/5,000; IP, dil: 1/500 | Proteintech | Cat#66008-3-Ig; RRID: AB_2749837 |
| Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L), WB, dil:1/50,000 | Jackson ImmunoResearch | Cat#Fs111-035-003; RRID: AB_2313567 |
| Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L), WB, dil:1/50,000 | Jackson ImmunoResearch | Cat#Fs115-035-003; RRID: AB_10015289 |
| Protein A/G PLUS-Agarose | Santa Cruz Biotechnology | Cat#sc-2003; RRID: AB_10201400 |
| Protein A+G Agarose | Beyotime | Cat#P2055 |
| Anti-Flag Affinity Gel | Bimake | Cat#B23101 |
| Pierce™ Protein A/G Magnetic Beads | Thermo Fisher Scientific | Cat#88803 |
| Anti-DYKDDDDK-Tag Mouse mAb (Agarose Conjugated) | Abmart | Cat#M20018 |
| Ni-NTA Agarose | Qiagen | Cat#30210 |
| Phospho-IRE1-S724 Rabbit mAb, B, dil:1/1,000 | Abclonal | Cat#AP0878; RRID: AB_2771207 |
| IRE1α (14C10) Rabbit mAb, WB, dil:1/1,000 | Cell Signaling Technology | Cat#3294S; RRID: AB_823545 |
| Phospho-eIF2α-S51 Rabbit mAb, WB, dil:1/1,000 | Abclonal | Cat#AP0692; RRID: AB_2863809 |
| eIF2α Antibody, WB, dil:1/1,000 | Cell Signaling Technology | Cat#9722S; RRID: AB_2230924 |
| Phospho-PERK (Thr980) (16F8) Rabbit mAb, WB, dil:1/1,000 | Cell Signaling Technology | Cat#3179S; RRID: AB_2095853 |
| PEK/PERK (Ab-981) Antibody, WB, dil:1/1,000 | Biorbyt | Cat#Ab-981; RRID: AB_10685785 |
| ATF4 (D4B8) Rabbit mAb, WB, dil:1/1,000 | Cell Signaling Technology | Cat#11815S; RRID: AB_2095853 |
| Anti-Ki67 antibody, IF, dil:1/100 | Abcam | Cat#ab16667; RRID: AB_302459 |
| Anti-NDUFB8 antibody, IF, dil:1/500 | Abcam | Cat#ab192878; RRID: AB_2847808 |
| Anti-HMGCS2 antibody, IF, dil:1/500 | Abcam | Cat#ab137043; RRID: AB_2749817 |
| Bacterial and virus strains | ||
| Adenoviral ceruplasmin shRNA | This paper | N/A |
| Adenoviral shLacz | This paper | N/A |
| HBAAV2/9-TBG-EGFP | Hanbio Biotechnology (Shanghai) | HH20190118GKL-AVP01 |
| HBAAV2/9-TBG-Cre-EGFP | Hanbio Biotechnology (Shanghai) | HH20190118GKL-AVP01 |
| Adenoviral PPARE-Luc | This paper | N/A |
| Adenoviral HMGCS2-Luc | This paper | N/A |
| Adenoviral CPT1α-Luc | This paper | N/A |
| Biological samples | ||
| Human liver tissue sample | Shanghai Jiao Tong University Affiliated Sixth People’s Hospital |
N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Oil Red O | Sigma-Aldrich | Cat#O0625 |
| Dexamethasone | Sigma-Aldrich | Cat#D4902 |
| Bovine Serum Albumin (BSA) | Sigma-Aldrich | Cat#A8806 |
| Protease Inhibitor Cocktail | Sigma-Aldrich | Cat#S8830 |
| PhosSTOP™ | Sigma-Aldrich | Cat#4906837001 |
| Sodium palmitate | Sigma-Aldrich | Cat#P9767 |
| Insulin | Roche | Cat#11376497001 |
| D-(+)-Glucose | Sigma-Aldrich | Cat#G7021 |
| Bathocuproinedisulfonic acid disodium salt (BCS) | Sigma-Aldrich | Cat#B1125 |
| Compound C (CC) | MedChemExpress | Cat#HY-13418A |
| Deferasirox (DFX) | Selleck | Cat#S1712 |
| Ammonium tetrathiomolybdate (TM) | Sigma-Aldrich | Cat#323446 |
| SP 600125 | Cayman Chemical | Cat#10010466 |
| Ex527 | Cayman Chemical | Cat#10009798 |
| GW 6471 | Cayman Chemical | Cat#11697 |
| GW 9578 | Cayman Chemical | Cat#10011211 |
| H89 | MedChemExpress | Cat#HY-15979 |
| Deferoxamine mesylate | Selleck | Cat#S5742 |
| Pyridoxal isonicotinoyl hydrazone | Cayman Chemical | Cat#19357 |
| Copper (II) sulfate | SHANGHAI LINFENG CHEMICAL | Cat#20160728 |
| SDS-PAGE Loading Buffer (non-Reducing, 5x) | Cwbiotech | Cat#CW0028S |
| Tetrakis(acetonitrile)copper(I) Hexafluorophosphate | Tcichemical | Cat#T2665 |
| D-isoascorbic acid | Sigma-Aldrich | Cat#856061 |
| IPTG | Yeasen | Cat#10902ES08 |
| Proteinase K Solution | ThermoFisher Scientific | Cat#AM2548 |
| Passive Lysis Buffer, 5X | Promega | Cat#E1941 |
| Penicillin/Streptomycin | ThermoFisher Scientific | Cat#15140-122 |
| Dulbecco’s modified Eagle’s medium (DMEM) | ThermoFisher Scientific | Cat#C11995500BT |
| ITS Liquid Media Supplement (100x) | Sigma-Aldrich | Cat# 41400045 |
| Medium 199 | Sigma-Aldrich | Cat#M2154 |
| L-Glutamine | Sigma-Aldrich | Cat#G3126 |
| Collagenase type I | Worthington biochemical corporation | Cat#4196 |
| Rat Collagen I | R&D Systems | Cat#3440-005-01 |
| PERCOLL | BIODEE | Cat#DE-17-0891-02G |
| RPMI 1640 medium | ThermoFisher Scientific | Cat#A10491-01 |
| DMEM/F12 | ThermoFisher Scientific | Cat#21331020 |
| TRIzol reagent | ThermoFisher Scientific | Cat#15596 |
| Lipofectamine RNAiMAX Transfection Reagent | ThermoFisher Scientific | Cat#13778075 |
| Fetal Bovine Serum | ThermoFisher Scientific | Cat#16000-044 |
| STZ | Sigma-Aldrich | Cat#18883-66-4 |
| Critical commercial assays | ||
| BCA assay kit | Thermo Fisher Scientific | Cat#23225 |
| Hiscript II Q RT SuperMix for qPCR | Vazyme, China | Cat#R222-01 |
| AceQ Universal SYBR qPCR Master Mix | Vazyme, China | Cat#Q511-02 |
| VAHTS mRNA-seq V2 Library Prep Kit for Illumina | Vazyme, China | Cat#NR601 |
| MitoTracker Red probe | ThermoFisher Scientific | Cat#M7512 |
| XF Cell Mito Stress Test Kit | Agilent Technologies | Cat#103015-100 |
| TG Assay kit | Applygene | Cat#E1013-105 |
| KOD -Plus Mutagenesis Kit | TOYOBO | Cat#SMK-101 |
| QIAquick PCR Purification Kit | Qiagen | Cat#28106 |
| Dual-Luciferase® Reporter Assay System | Promega | Cat#E1960 |
| DeadEnd™ Fluorometric TUNEL System | Promega | Cat#G3250 |
| NE-PER kit | Thermo Fisher | 78833 |
| AST kit | NJJCBIO | C010-2-1 |
| ALT kit | NJJCBIO | C009-2-1 |
| Deposited data | ||
| RNA sequencing data | This paper | Mendeley data (https://doi.org/10.17632/g5pxd2dcvm.1) |
| Experimental models: Cell lines | ||
| Mouse: AML12 cells | ATCC | Cat#CRL-2254 |
| Mouse: Primary hepatocytes | This paper | N/A |
| Human: L02 cells | Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. | N/A |
| Human: 293A | Invitrogen | Cat#R70507 |
| Human: HEK293T | ATCC | Cat#CRL-1573 |
| Experimental models: Organisms/strains | ||
| Mouse: C57BL/6J | GemPharmatech | Cat#N000013 |
| Mouse: ob/ob (B6/JGpt-Lepem1Cd25/Gpt) | GemPharmatech | Cat#T001461 |
| LDKO mice | GemPharmatech | N/A |
| Mouse: Cpflox/flox | Institute of Laboratory Animals Science | N/A |
| Mouse: global Cp KO | Institute of Laboratory Animals Science | N/A |
| Mouse: Albumin-Cre transgenic mice | The Jackson Laboratory | N/A |
| Mouse: AMPKα1flox/flox and α2flox/flox (AMPKαf/f) | Gift from Jingya Li (Chinese Academy of Sciences) | N/A |
| Mouse: AMPKα1 and α2 double liver KO (AMPKα-DKO) | Gift from Jingya Li (Chinese Academy of Sciences) | N/A |
| Oligonucleotides | ||
| Primers for qPCR, see Table S3 | This paper | N/A |
| Primers for plasmid construction, see Table S3 | This paper | N/A |
| siRNAs, see Table S4 | This paper | N/A |
| Recombinant DNA | ||
| pET28a-SCO1-HIS | This paper | N/A |
| SCO1-Flag | This paper | N/A |
| SCO1-His-Trunated | This paper | N/A |
| SCO1-C152/156A | This paper | N/A |
| pcDNA-3.0-PGC1α-Flag | This paper | N/A |
| CP-pLVX-IRES-Puro | This paper | N/A |
| pcDNA-3.1-AMPKα1-Myc | This paper | N/A |
| pcDNA-3.1-AMPKα1-Flag | Gift from Shengcai Lin (Xiamen University) | N/A |
| LKB1-Flag | Gift from Shengcai Lin (Xiamen University) | N/A |
| Software and algorithms | ||
| GraphPad Prism version 9.0 | GraphPad Software | https://www.graphpad.com/ |
| ImageJ | National Institutes of Health | https://imagej.nih.gov/ij/ |
| SPSS version 25.0 | IBM | https://www.ibm.com/analytics/spss-statistics-software |
| Other | ||
| High-fat diet | Research diet | Cat#D12492 |
| Chow diet | SLACOM | Cat#P1101F-25 |