Mass Spectrometry:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
Imaging:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
RNA sequencing:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
Chromatin Immunoprecipitation:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
Diagnostic Assay:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
Gene Expression:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
Recombinant:Article Title: Functional nutrient-genetic profiling reveals biotin and FBXW7 are essential to bypass glutamine addiction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Diethyl succinate Sigma-Aldrich Cat# 112402 2-ketobutyric acid Sigma-Aldrich Cat# K401 Dimethyl 2-oxoglutarate Sigma-Aldrich Cat# 349631 Thymidine Sigma-Aldrich Cat# T1895 L-aspartic acid Sigma-Aldrich Cat# A8949 L-asparagine monohydrate Sigma-Aldrich Cat# A4284 Uridine Sigma-Aldrich Cat# U3003 Biotin Sigma-Aldrich Cat# B4639 β-mercaptoethanol PAN-Biotech Cat# P07-05020 Puromycin Invivogen Cat# ant-pr-1 Blasticidin Sigma-Aldrich Cat# SBR00022 4-hydroxytamoxifen MedChemExpress Cat# HY-16950 UK5099 MedChemExpress Cat# HY-15475 Sulfasalazin Sigma-Aldrich Cat# S0883 Bafilomycin MedChemExpress Cat# HY-100558 PFI7 Structural Genomics Consortium Free sample ISRIB Sigma-Aldrich Cat# SML0843 Actinomycin D Sigma-Aldrich Cat# A9415 Polybrene Sigma-Aldrich Cat# TR-1003G Murine IL-2 recombinant protein, PeproTech® ThermoFisher (Gibco) Cat# 212-12 cOmpleteTM ULTRA protease inhibitor tablets Roche Cat# 05892970001 PierceTM Universal Nuclease Life Technologies Cat# 88702 Sodium deoxycholate Sigma-Aldrich Cat# D6750 IGEPAL® CA-630 (NP40 analog) Sigma-Aldrich Cat# I8896 TRI reagent Sigma-Aldrich Cat# T3809 M-MLV Reverse Transcriptase Sigma-Aldrich Cat# M1302-40KU RnaseOut Recombinant RNase Inhibitor Invitrogen Cat# 10777019 Random hexamers ThermoFisher Cat# N8080127 RNAse A Takara Bio Cat# 740505 Proteinase K Sigma-Aldrich Cat# SRE0005 Phenol-chloroform-isoamyl alcohol Sigma-Aldrich Cat# 77617 IRDye® 680RD Streptavidin LICORbio Cat# 926-68079 Critical commercial assays NucleoSpin Blood kit Machery Nagel Cat# 740954.20 DC Protein Assay Bio-Rad Cat# 5000111 Pierce High pH Reversed-Phase Peptide Fractionation Kit ThermoFisher Cat# 84868 PM-M1 BioLog Cat# 13101 PM-M2 BioLog Cat# 13102 PM-M3 BioLog Cat# 13103 PM-M4 BioLog Cat# 13104 BioLog MA dye BioLog Cat# 74351 PrestoBlueTM Cell Viability Reagent Life Technologies Cat# A13262 CellTiter-Glo® 2.0 Cell Viability Assay Promega Cat# G9242 CD8a + T cell isolation kit, mouse Miltenyi Biotec Cat# 130-104-075 (Continued on next page) e2 Molecular Cell 86, 901–916.e1–e10, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Deposited data 13 C- and 15 N-glutamine isotopic profiling in K562; 13 C-pyruvate isotopic profiling in primary CD8 + T cells This study MTBLS13799 13 C-pyruvate isotopic profiling in K562 This study 14002048 Multiple pathway targeted analysis of polar metabolites This study 14002048 Mass spectrometry data This study PXD057389 Spheroid imaging data This study https://www.doi.org/10.17632/328g7zyt9d.1 K562 RNAseq Slavoff et al. 81 GSE34740 Transcription factor ChIP-seq in K562 ENCODE 68 MYC(ENCFF801QJW), MNT(ENCFF611RGV), SIN3A(ENCFF085NLG) H3K27ac ChIP-seq in K562 ENCODE 68 ENCFF381NDD Experimental models: Cell lines Human: K562 American Type Culture Collection (ATCC) Cat# CCL-243; RRID: CVCL_0004 Human: HEK293T ATCC Cat# CRL-3216; RRID: CVCL_0063 Human: HeLa ATCC Cat# CCL-2; RRID: CVCL_0030 Human: MCF-7 ATCC Cat# HTB-22; RRID: CVCL_0031 Human: SW480 ATCC Cat# CCL-228; RRID: CVCL_0546 Human: UACC-257 Frederick Cancer Division of Cancer Treatment and Diagnostic (DCTD) RRID: CVCL_1779 Human: U-2 OS ATCC Cat# HTB-96; RRID: CVCL_0042 Human: A375 ATCC Cat# CRL-1619; RRID: CVCL_0132 Mouse: MC38 Sigma-Aldrich Cat# SCC172; RRID: CVCL_B288 Mouse: B16-F10 ATCC Cat# CRL-6475; RRID: CVCL_0159 Mouse: MycER mouse embryonic fibroblast Wise et al. 9 ; Linder et al. 19 N/A Oligonucleotides sgRNAs, see Table S9 Integrated DNA Technologies N/A ChIP primers, see Table S9 Integrated DNA Technologies N/A Random hexamers ThermoFisher Cat# N8080127 TaqMan Gene Expression Assay, human PC, FAM-MGB Life Technologies Cat# Hs01085875_g1 Taqman Gene Expression Assay, human TBP, VIC-MGB Life Technologies Cat# Hs00427620_m1 Recombinant DNA pLX304-GFP-V5 Kim et al. 82 Addgene, Cat# 193687; RRID: Addgene_193687 pLX304-PC Wiemann et al. 83 DNAsu, Cat# HsCD00436386 pWPI-Neo-GFP-3xFLAG Jourdain et al. 84 Addgene, Cat# 201639; RRID: Addgene_201639 pWPI-SLC7A11-HA Jourdain et al. 84 Addgene, Cat# 201643; RRID: Addgene_201643 pMXS-SLC1A3 Birsoy et al. 30 Addgene, Cat# 72873; RRID: Addgene_72873 pLVX-TetOne-Puro-GFP-3xFLAG This study N/A pLVX-TetOne-Puro-LbNOX Titov et al. 31 and Choe et al. 85 Addgene, Cat# 234984; RRID: Addgene_234984 pTwist-Lenti-CMV-BSD-GFP-3xFLAG This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-WT This study Twist Bioscience pTwist-Lenti-CMV-BSD-PC-K1144R This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-WT This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R465C This study Twist Bioscience (Continued on next page) Molecular Cell 86, 901–916.e1–e10, March 5, 2026 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER pTwist-Lenti-CMV-Puro-FBXW7-R505C This study Twist Bioscience pTwist-Lenti-CMV-Puro-FBXW7-R479Q This study Twist Bioscience psPAX2 Didier Trono Addgene, Cat# 12260; RRID: Addgene_12260 pMD2.G Didier Trono Addgene, Cat# 12259; RRID: Addgene_12259 lentiCRISPR v2 Sanjana et al. 86 Addgene, Cat# 52961; RRID: Addgene_52961 Human sgRNA library Brunello (lentiCRISPR v2) Sanson et al. 87 Addgene, Cat# 73179; RRID: Addgene_73179 Software and algorithms Prism GraphPad Software https://www.graphpad.com/ Fiji Schindelin et al. 88 https://imagej.net/software/fiji/ IGV Robinson et al. 89 https://igv.org/ GSEA Subramanian et al. 90 https://www.gsea-msigdb.org/gsea/index.jsp MetaboAnalyst Pang et al. 29 https://www.metaboanalyst.ca/ Spectronaut Biognosys Schlieren https://biognosys.com/software/spectronaut/ IDPicker algorithm Ma et al. 91 https://proteowizard.sourceforge.io/idpicker/ MaxLFQ algorithm Cox et al. 92 https://www.maxquant.org/ Perseus Tyanova et al. 93 https://maxquant.net/perseus/ MassHunter Quantitative Analysis Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software ProFinder Agilent Technologies https://www.agilent.com/en/product/ software-informatics/massspectrometry-software/dataanalysis/mass-profilerprofessional-software R R Core Team 94 http://cran.r-project.org/ LOWESS/Spline normalization algorithm Tsugawa et al. 95 http://prime.psc.riken.jp Xcalibur ThermoFisher https://www.thermofisher.com/us/en/ home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ lc-ms-data-acquisition-software/ xcalibur-data-acquisitioninterpretation-software.html Compound Discoverer ThermoFisher https://www.thermofisher.com/ch/ en/home/industrial/mass-spectrometry/ liquid-chromatography-massspectrometry-lc-ms/lc-ms-software/ multi-omics-data-analysis/compounddiscoverer-software.html?cid= E.25CMD.CD103.25418.01 Skyline University of Washington https://skyline.ms/home/software/ Skyline/project-begin.view Other U-bottom NunclonTM SpheraTM 96-Well plates ThermoFisher Cat# 174925 NuncTM MicroWellTM 96-Well plates Nunc Cat# 137101 8-16% NovexTM WedgeWellTM Tris-Glycine gels ThermoFisher Cat# XP08160BOX Dynabeads Protein G ThermoFisher Cat# 10007D Dynabeads Protein A ThermoFisher Cat# 10001D e4 Molecular Cell 86, 901–916.e1–e10, March 5, 2026
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