Gel Extraction:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
Plasmid Preparation:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
Purification:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
Enzyme-linked Immunosorbent Assay:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
Cell Isolation:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
SYBR Green Assay:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
Cell Stimulation:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
Lactate Assay:Article Title: Metabolic reinvigoration of NK cells by IL-21 enhances immunotherapy against MHC class I-deficient solid tumors.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Halt TM Protease Inhibitor Cocktail Thermo Scientific TM Cat # 78429 BshTI (AgeI) Thermo Scientific TM Cat # ER1461 FastDigest EcoRI Thermo Scientific TM Cat # FD0275 T4 DNA Ligase Thermo Scientific TM Cat # EL0016 Bp2 (BbsI) Thermo Scientific TM Cat # ER1011 Recombinant Mouse IL-2 Biolegend Cat # 575408 Brefeldin A Solution (1,000 x) Biolegend Cat # 420601 Sodium citrate Sigma-Aldrich CAS 6132-04-3 Sodium chloride Sigma-Aldrich CAS 7647-14-5 Tris–HCl Sigma-Aldrich CAS 1185-53-1 Acetic acid, 99.5%, pure Sigma-Aldrich CAS 64-19-7 2-NBDG Sigma-Aldrich CAS 186689-07-6 Oligomycin Sigma-Aldrich CAS 1404-19-9 Carbonyl cyanide 4- (trifluoromethoxy)phenylhydrazone(FCCP) Sigma-Aldrich CAS 370-86-5 Rotenone Sigma-Aldrich CAS 83-79-4 Antimycin A Sigma-Aldrich CAS 1397-94-0 2-Deoxy-D-glucose = 98% (GC), crystalline Sigma-Aldrich CAS 154-17-6 Ammonium persulfate Sigma-Aldrich CAS 7727-54-0 Bovine Serum Albumin Sigma-Aldrich Cat # A-3059 2-mercaptoethanol Sigma-Aldrich CAS 60-24-2 Non Fat Dry Milk Cell Signaling Technology Cat # 9999 Cell Lysis Buffer (10 x) Cell Signaling Technology Cat # 9803 Methanol Fisher Chemical CAS 67-56-1 Ethanol Fisher Chemical CAS 64-17-5 Bolt TM LDS Sample Buffer (4 x) Invitrogen TM Cat #B0008 Trypan Blue Stain (0.4%) for use with the Countess TM Automated Cell Counter Invitrogen TM Cat #T10282 S.O.C. .. Medium Invitrogen TM Cat # 15544034 EDTA pH 8.0 (0.5M) Invitrogen TM Cat # 15575020 Tris/Tricine/SDS Buffer (10 x) Bio-Rad Cat # 1610744 Tris/Glycine Buffer (10 x) Bio-Rad Cat # 1610734 40% Acrylamide/Bis Solution, 29:1 Bio-Rad Cat # 1610146 STF-31 MedChemExpress CAS 724741-75-7 Lactate Dehydrogenase A Inhibitor, FX11 EMD Millipore CAS 213971-34-7 Critical commercial assays GeneJET Gel Extraction and DNA Cleanup Micro Kit Thermo Scientific TM Cat #K0831 GeneJET Plasmid Miniprep Kit Thermo Scientific TM Cat #K0502 PureLink TM Fast Low-Endotoxin Maxi Plasmid Purification Kit Thermo Scientific TM Cat # A36228 Mouse IL-21 DuoSet ELISA R&D Cat # DY594-05 Zombie Aqua TM Fixable Viability Kit Biolegend Cat # 423102 NK Cell Isolation Kit, mouse Miltenyi Biotec Cat # 130-115-818 PowerUp SYBR Green Master Mix Appliedbiosysterms Cat # 01034944 PageRuler TM Prestained Protein Ladder Thermo Scientific TM Cat # 26616 eBioscience TM Cell Stimulation Cocktail Invitrogen TM Cat # 00-4970-03 Lactate Assay kit (C/F) BioVision Cat #K607 MycoAlert® Mycoplasma Detection Kit Lonza Cat # LT07-318 (Continued on next page) Cell Reports 45, 117035, March 24, 2026 21 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data Bulk RNA-seq data This paper GEO: GSE28973 scRNA-seq data This paper GEO: GSE282121 Oligonucleotides (5 ′ to 3 ′ ) sgLdha-1-TTGGTGAGTATACCTTGGGC This paper N/A sgLdha-2-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-3-GGGACTTGGTGAGTATACCT This paper N/A sgLdha-4-GCACCGGCTAAAGTCACTGT This paper N/A sgScramble-TGCGAATACGCCCACGCGAT This paper N/A Recombinant DNA Plasmid gWIZ_Kozak_mIgGSP_mouse IL-21− Fc This paper N/A Plasmid plenti guide Thy1.1 Addgene RRID: Addgene_128063 Plasmid MSCV puro Addgene RRID: Addgene_68469 Plasmid pCMV-VSV-G Addgene RRID: Addgene_8454 Plasmid pCMV-dR8.2 dvpr Addgene RRID: Addgene_8455 Experimental models: Cell lines B16F10_β2m − /− Gift from Prof. David H. Raulet N/A CT26_β2m − /− Gift from Prof. David H. Raulet N/A RMA-S Gift from Prof. Werner Held N/A Phoenix-Eco ATCC Cat # CRL-3214 TM , RRID: CVCL_H717 MDA-MB-231 ATCC Cat # HTB-26 TM RRID: CVCL_0062 U87 ATCC Cat # HTB-14 TM RRID: CVCL_0022 HepG2 Haixing Biosciences Cat # TCH-C196 HCT116 Haixing Biosciences Cat # TCH-C185 Experimental models: Organisms/strains Mouse: C57BL/6J Charles River Laboratories Strain #: 000664 RRID: IMSR_JAX:000664 Mouse: BALB/cByJ Charles River Laboratories Strain #: 001026 RRID: IMSR_JAX:001026 Mouse: B6.SJL-Ptprc a Pepc b /BoyCrl The Jackson Laboratory Strain #: 002014 RRID: IMSR_JAX:002014 Mouse: Gt(ROSA)26Sor tm1.1(CAG-cas9 * ,− EGFP)Fezh /J The Jackson Laboratory Strain #: 024858 RRID: IMSR_JAX:026179 Mouse: NOD-Prkdcscid Il2rgem1/Smoc (M-NSG) Shanghai Model Organisms Center, Inc. Cat NO. NM-NSG-001 RRID: IMSR_NM-NSG-001 Software and algorithms GraphPad Prism 10 https://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 https://www.flowjo.com/flowjo/ overview RRID:SCR_008520 Rstudio v.4.3.1 https://cran.rstudio.com/bin/ windows/base/old/4.3.1/ RRID:SCR_000432 featureCounts Subread package N/A Agilent Masshunter Quantitative Analysis software Version B.07.00 http://www.agilent.com/en-us/ products/software-informatics/ masshunter-suite/masshunter/ masshunter-software RRID:SCR_015040 MRM PROBS Tsugawa et al. 79 N/A Other Amicon® Ultra-15 Centrifugal Filter Unit Merck Millipore Cat # UFC901024 Millex-HV Syringe Filter Unit, 0.22 μm Merck Millipore Cat # SLGSV255F (Continued on next page) 22 Cell Reports 45, 117035, March 24, 2026
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