KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-PHGDH polyclonal antibody produced in rabbit | Sigma-Aldrich | Cat# HPA021241; RRID: AB_1855299 |
| Anti-SREBP-1 (2A4) monoclonal antibody produced in mouse | Santa Cruz Biotechnology | Cat# sc-13551; RRID: AB_628282 |
| Anti-mTOR antibody produced in rabbit, unconjugated | Cell Signaling Technology | Cat# 2972; RRID: AB_330978 |
| Anti-Phospho-p70 S6 Kinase (Thr389) polyclonal antibody produced in rabbit | Cell Signaling Technology | Cat# 9205; RRID: AB_330944 |
| Anti-p70 S6 Kinase polyclonal antibody produced in rabbit | Cell Signaling Technology | Cat# 9202; RRID: AB_331676 |
| Anti-ITFG2 (F-11) monoclonal antibody produced in mouse, conjugated to AlexaFluor 488 | Santa Cruz Biotechnology | Cat# sc-271420; RRID: AB_10611069 |
| Anti-NPRL2 (F-3) monoclonal antibody produced in mouse, conjugated to AlexaFluor 488 | Santa Cruz Biotechnology | Cat# sc-376986 |
| Peroxidase IgG Fraction Monoclonal Mouse Anti-Rabbit IgG, light chain specific antibody | Jackson ImmunoResearch Labs | Cat# 211-032-171; RRID: AB_2339149 |
| Bacterial and virus strains | ||
| PHGDH MISSION shRNA Bacterial Glycerol Stock | Sigma-Aldrich | SHCLNG-NM_006623 TRCN0000028520 |
| PHGDH MISSION shRNA Bacterial Glycerol Stock | Sigma-Aldrich | SHCLNG-NM_006623 TRCN0000233033 |
| MISSION® pLKO.1-puro TurboGFP shRNA Control Transduction Particles | Sigma-Aldrich | SHC004V |
| Incucyte® NucLight Red Lentivirus Reagent (EF-1 Alpha Promoter, Puromycin selection) | Sartorius | Cat# 4476 |
| Biological samples | ||
| Osteosarcoma tumor microarray | University of Texas MD Anderson Cancer Center | University of Texas MD Anderson Cancer Center |
| Chemicals, peptides, and recombinant proteins | ||
| Alfa Aesar L-Serine, Cell Culture Reagent | Fisher Scientific | Cat# AAJ6218709 |
| Glycine 1 M solution | Sigma-Aldrich | Cat# 67419 |
| Methotrexate | Cayman Chemical | Cat# 13960 |
| NCT-503 Inactive Control | Sigma-Aldrich | Cat# SML1671 |
| NCT-503 | Sigma-Aldrich | Cat# SML1659 |
| PKUMDL-WQ-2101 | Sigma-Aldrich | Cat# SML1970 |
| BPTES | Sigma-Aldrich | Cat# SML0601 |
| Rapamycin | Sigma-Aldrich | Cat# R8781 |
| Perhexiline maleate salt | Sigma-Aldrich | Cat# SML0120 |
| InSolution Etomoxir - CAS 828934-41-4 - Calbiochem | Sigma-Aldrich | Cat# 5.09455 |
| Fetal Bovine Serum, dialyzed, US origin, One Shot format | GIBCO, Thermo Fisher Scientific | Cat# A3382001 |
| Lipoprotein Depleted Fetal Bovine Serum | Kalen Biomedical | Cat# 880100-5 |
| YOYO-1 Iodide (491/509) −1 mM Solution in DMSO | Invitrogen | Cat# Y3601 |
| Palmitic acid | Sigma-Aldrich | Cat# P0500-25G |
| D-Glucose (U-13C6, 99%) | Cambridge Isotope Laboratories | Cat# CLM-1396-0.5 |
| Palmitic Acid (U-13C16, 98%) | Cambridge Isotope Laboratories | Cat# CLM-409-PK |
| BODIPY 493/503 (4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene) | Invitrogen | Cat# D3922 |
| Polyethylene glycol 300 | Sigma-Aldrich | Cat# 8.17002 |
| (2-Hydroxypropyl)-β-cyclodextrin | Sigma-Aldrich | Cat# H107 |
| LysoTracker Deep Red | Invitrogen | Cat# L12492 |
| Critical commercial assays | ||
| Quick Start Bradford Protein Assay Kit 2 | Bio-Rad | Cat# 5000202 |
| Seahorse XF Cell Mito Stress Test Kit | Agilent | Cat# 103015-100 |
| Seahorse XF Glycolytic Rate Test Kit | Agilent | Cat# 103710-100 |
| Seahorse XF Mito Fuel Flex Test Kit | Agilent | Cat# 103260-100 |
| Direct-zol RNA Miniprep Plus Kit | Zymo Research | Cat# R2070 |
| MILLIPLEX MAP Total Akt/mTOR Magnetic Bead 11-Plex Kit - Cell Signaling Multiplex Assay | Millipore | Cat# 48-612MAG |
| Phosphoglycerate Dehydrogenase (PHGDH) Activity Assay Kit | PromoCell | Cat# PK-CA577-K569 |
| PicoProbe Lactate Fluorometric Assay Kit | BioVision | Cat# K638 |
| Alpha Ketoglutarate (alpha KG) Assay Kit | Abcam | Cat# ab83431 |
| Experimental models: cell lines | ||
| NOS1 | RIKEN BioResource Research Center | Cat# RCB1032; RRID: CVCL_1610 |
| Saos2 | ATCC | Cat# HTB-85; RRID: CVCL_0548 |
| U2OS | ATCC | Cat# HTB-96; RRID: CVCL_0042 |
| MG63 | ATCC | Cat# CRL-1427; RRID: CVCL_0426 |
| MNNG | ATCC | Cat# CRL-1547; RRID: CVCL_0439 |
| MDA-MB-231 | ATCC | Cat# CRL-12532; RRID: CVCL_0062 |
| MDA-MB-468 | ATCC | Cat# HTB-132; RRID: CVCL_0419 |
| Experimental models: organisms/strains | ||
| Mouse: athymic nude mice (female, 4–6 weeks old, homozygous for Foxn1nu) | The Jackson Laboratory | RRID: IMSR_JAX:002019 |
| Software and algorithms | ||
| GraphPad Prism | GraphPad | RRID: SCR_002798 |
| Incucyte® Base Analysis Software | Sartorius | https://www.essenbioscience.com/en/products/software/incucyte-base-software/ |
| IncuCyte® S3 Software | Sartorius | https://www.essenbioscience.com/en/products/software/incucyte-s3-software-v2018b/ |
| CalcuSyn | Biosoft | http://www.biosoft.com/w/calcusyn.htm |
| Compass for SimpleWestern | proteinsimple | https://www.proteinsimple.com/software_compass_simplewestern.html |
| Seahorse Wave | Agilent Technologies | RRID: SCR_014526 |
| MetaboAnalyst 4.0 Software | MetaboAnalyst | https://www.metaboanalyst.ca/ |