Saline:Article Title: A glutamine metabolism-related gene signature for liver fibrosis prediction integrating single-cell and bulk transcriptomic analysis with multi-cohort validation
Article Snippet: Equal amounts of protein (30 μg per lane) were separated by SDS-PAGE on 10% polyacrylamide gels and transferred onto PVDF membranes (Millipore, USA). .. Membranes were blocked with 5% non-fat milk in TBST (Tris-buffered saline with 0.1% Tween-20) at room temperature for 1 hour, then incubated overnight at 4°C with primary antibodies: anti-GOT1 (Proteintech, Cat No. 14886-1-AP, diluted 1:1000), anti-GLS (Proteintech, Cat No. 12855-1-AP, diluted 1:5000), and anti-β-actin (Abiowell, Cat No. AWA80113, diluted 1:5000). ..
Incubation:Article Title: A glutamine metabolism-related gene signature for liver fibrosis prediction integrating single-cell and bulk transcriptomic analysis with multi-cohort validation
Article Snippet: Equal amounts of protein (30 μg per lane) were separated by SDS-PAGE on 10% polyacrylamide gels and transferred onto PVDF membranes (Millipore, USA). .. Membranes were blocked with 5% non-fat milk in TBST (Tris-buffered saline with 0.1% Tween-20) at room temperature for 1 hour, then incubated overnight at 4°C with primary antibodies: anti-GOT1 (Proteintech, Cat No. 14886-1-AP, diluted 1:1000), anti-GLS (Proteintech, Cat No. 12855-1-AP, diluted 1:5000), and anti-β-actin (Abiowell, Cat No. AWA80113, diluted 1:5000). ..
Article Title: Radiotherapy followed by metformin neoadjuvant therapy drives superior pancreatic cancer ferroptosis and augments anti-tumor immunity
Article Snippet: .. After deparaffinization nd antigen retrieval in citrate buffer, sections were permeabilized with .3% Triton X-100, blocked with 5% BSA, and incubated overnight at °C with primary antibodies: anti-rabbit Ki-67 (Proteintech, #28074-1- P) and anti-mouse GOT1 (Proteintech, #60317-1-Ig). ..
Article Title: Radiotherapy followed by metformin neoadjuvant therapy drives superior pancreatic cancer ferroptosis and augments anti-tumor immunity
Article Snippet: .. Sections were subsequently dewaxed and then incubated ith anti-human GOT1 (Proteintech, #14886-1-AP), anti-mouse GOT1 Proteintech, #60317-1-Ig), and anti-mouse CD8 (Proteintech, #66868- -Ig), and anti-mouse CD11c (Abclonal, #A1508), respectively, at °C overnight. .. After being stained with hematoxylin, the sections were s c h a 0 4 A A l r 2 S 1 t 2 f s d i e t b f P v 2 s w y S t t 0 a s 3 3 a e c p c t M t c e ( p a b r r i o [ l P c t a R e c 1 o m & ( R t R i ( P t m c e t a w c w t t f 3 n t s p i c y a m f ( M t p p i a i P a p c e ealed with neutral resin, scanned and analyzed under an Olympus miroscope.
other:Article Title: Aspartate availability drives differential engagement of the malate-aspartate shuttle.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA lentiCRISPR v2 Sanjana et al. 69 Addgene Plasmid #52961 pUC57-LbNOX Titov et al. 29 Addgene Plasmid #75285 N174-MCS Adam Karpf Addgene Plasmid #81061 GpASNase1 Sullivan et al. 38 Addgene Plasmid #121526 pCW57-GFP-P2A-MCS (Neo) Barger et al. 70 Addgene Plasmid #89181 psPAX2 Didier Trono Addgene Plasmid #12260 pMD2.G Didier Trono Addgene Plasmid #12259 pCW57-GFP-P2A-ByASPase This paper Addgene Plasmid #252094 pCW57-GFP-P2A-EcASPase This paper Addgene Plasmid #252079 Software and algorithms R Cran https://www.r-project.org Prism 9-10 Graphpad https://www.graphpad.com IsoCor v2.0 Millard et al. 71 N/A Quant Studio 5-6 Thermo Fisher Scientific https://www.thermofisher.com/us/en/ home/life-science/pcr/real-time-pcr/realtime-pcr-instruments/quantstudiosystems/features.html#design-andanalysis-software MassHunter Agilent https://www.agilent.com/en/product/ software-informatics/mass-spectrometrysoftware/data-analysis Wave Desktop Agilent https://www.agilent.com/en/product/cell- analysis/real-time-cell-metabolic-analysis/ xf-software/seahorse-wave-desktopsoftware-740897?srsltid=AfmBOoojwqu EuqNM6xcIjeIHptR1Aj2AEeiKGlha32Itn1iQsuyp1LH e3 Molecular Cell 86, 954–967.e1–e7, March 5, 2026
Recombinant:Article Title: Aspartate availability drives differential engagement of the malate-aspartate shuttle.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse IgG HRP Linked Whole Ab Sigma-Aldrich/Cytiva Cat#GENA931-1ML Rabbit IgG HRP Linked Whole Ab Sigma-Aldrich/Cytiva Cat#GENA934-1ML; RRID: AB_2722659 MDH1 Polyclonal antibody Proteintech Cat15904-1-AP; RRID: AB_2143279 SLC25A11 Polyclonal antibody Proteintech Cat#12253-1-AP; RRID: AB_2877840 GOT1 Polyclonal antibody Proteintech Cat#14886-1-AP; RRID: AB_2113630 ARALAR/AGC1 (D5I6I) Rabbit Monoclonal Antibody Cell Signaling Technology Cat#64169S; RRID: AB_2799655 SLC25A13 Polyclonal Antibody Invitrogen Cat#PA5-21991; RRID: AB_11153758 GOT2 Polyclonal antibody Proteintech Cat#14800-1-AP; RRID: AB_2247898 Anti-MDH2 antibody Abcam Cat#ab96193; RRID: AB_10679348 ANTI-FLAG M2 antibody, Mouse monoclonal Sigma-Aldrich Cat#F1804; RRID: AB_262044 Anti-Vinculin Antibody Sigma-Aldrich Cat#V9131; RRID: AB_477629 Chemicals, peptides, and recombinant proteins DMSO Sigma-Aldrich Cat#D2650-100ML Puromycin Dihydrochloride ThermoFisher Scientific/Gibco Cat#A1113803 G418 Sulfate Solution GeminiBio Cat#400-113 Doxycycline hyclate Sigma-Aldrich Cat#D9891 Hexadimethrine bromide (polybrene) Sigma-Aldrich Cat#107689 RPMI MSKCC media core N/A DMEM, high glucose MSKCC media core N/A DMEM:F-12 MSKCC media core N/A L-Glutamine MSKCC media core N/A RPMI, minus glucose minus glutamine MSKCC media core N/A DMEM, high glucose, minus serine, minus glycine MSKCC media core N/A BenchMarkTM Fetal Bovine Serum Gemini Bio Cat#100-106 Dialyzed Fetal Bovine Serum Gemini Bio Cat#100-108-500 DMEM, no glucose, no glutamine, no phenol red Gibco Cat#A1443001 D-(+)-Glucose solution 45% Corning Cat#45001-116 Horse Serum, heat inactivated, New Zealand origin ThermoFisher Scientific/Gibco Cat#26050070 RIPA Buffer (10x) Cell Signaling Technologies Cat#9806 3-Isobutyl-1-methylxanthine Sigma-Aldrich Cat#I5879 Dexamethasone Sigma-Aldrich Cat#D8893 Troglitazone Sigma-Aldrich Cat#T2573 Insulin human Sigma-Aldrich Cat#I3536 Linoleic Acid-Oleic Acid-Albumin, 100x Sigma-Aldrich Cat#L9655 Pyridine Thermo Fisher Scientific Cat#TS-27530 Methoxyamine hydrochloride Sigma-Aldrich Cat#226904 Thermo ScientificTM MSTFA Thermo Fisher Scientific Cat#PI48910 Methanol, OptimaTM for HPLC, Fisher ChemicalTM Fisher Scientific Cat#A454SK-4 (Continued on next page) e1 Molecular Cell 86, 954–967.e1–e7, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Ethyl acetate, ACS, 99.5+% Fisher Scientific Cat#AA31344M1 Nutlin-3a Selleckchem Cat#S8059 Etoposide Sigma-Aldrich Cat#E1383 Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) Sigma-Aldrich Cat#C2920 Sodium Pyruvate (100 mM) Gibco Cat#11360070 TRIzolTM Reagent Invitrogen Cat#15596018 Power SYBRTM Green PCR Master Mix Applied Biosystems Cat#4367659 L-Asparagine monohydrate Sigma-Aldrich Cat#A7094 L-Serine Sigma-Aldrich Cat#S4311 Glycine Sigma-Aldrich Cat#50046 L-Glutamic acid monosodium salt monohydrate Sigma-Aldrich Cat#49621 N-Acetyl-L-cysteine Sigma-Aldrich Cat#A9165 Oil Red O solution Sigma-Aldrich Cat#O1391 X-Gal Roche Cat#10651745001 D-Glucose (U-1 3 C6, 99%) Cambridge Isotopes Laboratories, Inc. Cat#CLM-1396-10 L-Glutamine (1 3 C5, 99%) Cambridge Isotopes Laboratories, Inc. Cat#CLM-1822-H-1 PierceTM ECL Western Blotting Substrate Thermo Fisher Scientific Cat#32106 Critical commercial assays PierceTM BCA Protein Assay Kits Thermo Fisher Scientific Cat#23225 Seahorse XF Cell Mito Stress Test Kit Agilent Cat#103015-100 iScriptTM cDNA Synthesis Kit Biorad Cat#1708891 QIAGEN Plasmid Maxi Kit Qiagen Cat#12163 QIAGEN Plasmid Mini Kit Qiagen Cat#12125 QIAquick PCR & Gel Cleanup Kit Qiagen Cat#28506 QIAEX II Gel Extraction Kit Qiagen Cat#20051 Deposited data DepMap Portal R23Q4 Broad; Arafeh et al. 65 https://depmap.org/portal Original uncropped western blots This paper Mendeley Data: https://doi.org/10.17632/ k43x8thkyn.2 Experimental models: Cell lines C3H/10T1/2, Clone 8 ATCC Cat#CCL-226 C2C12 ATCC Cat#CRL-1772 293T ATCC Cat#CRL-3216 A549 ATCC Cat#CRM-CCL-185 SW872 ATCC Cat#HTB-92 Calu1 ATCC Cat#HTB-54 NCI-H1975 ATCC Cat#CRL-5908 NCI-H2122 ATCC Cat#CRL-5985 NCI-H2170 ATCC Cat#CRL-5928 NCI-H1993 ATCC Cat#CRL-5909 Oligonucleotides Guide RNAs for metabolic enzymes (STAR Methods) This paper and Morris et al. 66 and Wang et al. 67 IDT qPCR primers (STAR Methods) This paper and Lercher et al. 68 IDT Codon optimized ASPases (STAR Methods) This paper Twist Bioscience (Continued on next page) Molecular Cell 86, 954–967.e1–e7, March 5, 2026 e2
High Performance Liquid Chromatography:Article Title: Aspartate availability drives differential engagement of the malate-aspartate shuttle.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse IgG HRP Linked Whole Ab Sigma-Aldrich/Cytiva Cat#GENA931-1ML Rabbit IgG HRP Linked Whole Ab Sigma-Aldrich/Cytiva Cat#GENA934-1ML; RRID: AB_2722659 MDH1 Polyclonal antibody Proteintech Cat15904-1-AP; RRID: AB_2143279 SLC25A11 Polyclonal antibody Proteintech Cat#12253-1-AP; RRID: AB_2877840 GOT1 Polyclonal antibody Proteintech Cat#14886-1-AP; RRID: AB_2113630 ARALAR/AGC1 (D5I6I) Rabbit Monoclonal Antibody Cell Signaling Technology Cat#64169S; RRID: AB_2799655 SLC25A13 Polyclonal Antibody Invitrogen Cat#PA5-21991; RRID: AB_11153758 GOT2 Polyclonal antibody Proteintech Cat#14800-1-AP; RRID: AB_2247898 Anti-MDH2 antibody Abcam Cat#ab96193; RRID: AB_10679348 ANTI-FLAG M2 antibody, Mouse monoclonal Sigma-Aldrich Cat#F1804; RRID: AB_262044 Anti-Vinculin Antibody Sigma-Aldrich Cat#V9131; RRID: AB_477629 Chemicals, peptides, and recombinant proteins DMSO Sigma-Aldrich Cat#D2650-100ML Puromycin Dihydrochloride ThermoFisher Scientific/Gibco Cat#A1113803 G418 Sulfate Solution GeminiBio Cat#400-113 Doxycycline hyclate Sigma-Aldrich Cat#D9891 Hexadimethrine bromide (polybrene) Sigma-Aldrich Cat#107689 RPMI MSKCC media core N/A DMEM, high glucose MSKCC media core N/A DMEM:F-12 MSKCC media core N/A L-Glutamine MSKCC media core N/A RPMI, minus glucose minus glutamine MSKCC media core N/A DMEM, high glucose, minus serine, minus glycine MSKCC media core N/A BenchMarkTM Fetal Bovine Serum Gemini Bio Cat#100-106 Dialyzed Fetal Bovine Serum Gemini Bio Cat#100-108-500 DMEM, no glucose, no glutamine, no phenol red Gibco Cat#A1443001 D-(+)-Glucose solution 45% Corning Cat#45001-116 Horse Serum, heat inactivated, New Zealand origin ThermoFisher Scientific/Gibco Cat#26050070 RIPA Buffer (10x) Cell Signaling Technologies Cat#9806 3-Isobutyl-1-methylxanthine Sigma-Aldrich Cat#I5879 Dexamethasone Sigma-Aldrich Cat#D8893 Troglitazone Sigma-Aldrich Cat#T2573 Insulin human Sigma-Aldrich Cat#I3536 Linoleic Acid-Oleic Acid-Albumin, 100x Sigma-Aldrich Cat#L9655 Pyridine Thermo Fisher Scientific Cat#TS-27530 Methoxyamine hydrochloride Sigma-Aldrich Cat#226904 Thermo ScientificTM MSTFA Thermo Fisher Scientific Cat#PI48910 Methanol, OptimaTM for HPLC, Fisher ChemicalTM Fisher Scientific Cat#A454SK-4 (Continued on next page) e1 Molecular Cell 86, 954–967.e1–e7, March 5, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER Ethyl acetate, ACS, 99.5+% Fisher Scientific Cat#AA31344M1 Nutlin-3a Selleckchem Cat#S8059 Etoposide Sigma-Aldrich Cat#E1383 Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) Sigma-Aldrich Cat#C2920 Sodium Pyruvate (100 mM) Gibco Cat#11360070 TRIzolTM Reagent Invitrogen Cat#15596018 Power SYBRTM Green PCR Master Mix Applied Biosystems Cat#4367659 L-Asparagine monohydrate Sigma-Aldrich Cat#A7094 L-Serine Sigma-Aldrich Cat#S4311 Glycine Sigma-Aldrich Cat#50046 L-Glutamic acid monosodium salt monohydrate Sigma-Aldrich Cat#49621 N-Acetyl-L-cysteine Sigma-Aldrich Cat#A9165 Oil Red O solution Sigma-Aldrich Cat#O1391 X-Gal Roche Cat#10651745001 D-Glucose (U-1 3 C6, 99%) Cambridge Isotopes Laboratories, Inc. Cat#CLM-1396-10 L-Glutamine (1 3 C5, 99%) Cambridge Isotopes Laboratories, Inc. Cat#CLM-1822-H-1 PierceTM ECL Western Blotting Substrate Thermo Fisher Scientific Cat#32106 Critical commercial assays PierceTM BCA Protein Assay Kits Thermo Fisher Scientific Cat#23225 Seahorse XF Cell Mito Stress Test Kit Agilent Cat#103015-100 iScriptTM cDNA Synthesis Kit Biorad Cat#1708891 QIAGEN Plasmid Maxi Kit Qiagen Cat#12163 QIAGEN Plasmid Mini Kit Qiagen Cat#12125 QIAquick PCR & Gel Cleanup Kit Qiagen Cat#28506 QIAEX II Gel Extraction Kit Qiagen Cat#20051 Deposited data DepMap Portal R23Q4 Broad; Arafeh et al. 65 https://depmap.org/portal Original uncropped western blots This paper Mendeley Data: https://doi.org/10.17632/ k43x8thkyn.2 Experimental models: Cell lines C3H/10T1/2, Clone 8 ATCC Cat#CCL-226 C2C12 ATCC Cat#CRL-1772 293T ATCC Cat#CRL-3216 A549 ATCC Cat#CRM-CCL-185 SW872 ATCC Cat#HTB-92 Calu1 ATCC Cat#HTB-54 NCI-H1975 ATCC Cat#CRL-5908 NCI-H2122 ATCC Cat#CRL-5985 NCI-H2170 ATCC Cat#CRL-5928 NCI-H1993 ATCC Cat#CRL-5909 Oligonucleotides Guide RNAs for metabolic enzymes (STAR Methods) This paper and Morris et al. 66 and Wang et al. 67 IDT qPCR primers (STAR Methods) This paper and Lercher et al. 68 IDT Codon optimized ASPases (STAR Methods) This paper Twist Bioscience (Continued on next page) Molecular Cell 86, 954–967.e1–e7, March 5, 2026 e2
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