|
Antibodies
|
| α-mouse Gr1 Biotin |
Tonbo Biosciences |
Clone RB6–8C5; RRID:AB_2621652 |
| α-mouse Gr1 FITC |
Tonbo Biosciences |
Clone RB6–8C5; RRID:AB_2621721 |
| α-mouse Gr1 PE-Cy7 |
Tonbo Biosciences |
Clone RB6–8C5; RRID:AB_2621870 |
| α-mouse Gr1 APC |
Tonbo Biosciences |
Clone RB6–8C5; RRID:AB_2621610 |
| α-mouse Gr1 PE |
Tonbo Biosciences |
Clone RB6–8C5; RRID:AB_2621803 |
| α-mouse CD2 Biotin |
Biolegend |
Clone RM2–5; RRID:AB_312650 |
| α-mouse CD2 FITC |
Tonbo Biosciences |
Clone RM2–5; RRID:AB_2621657 |
| α-mouse CD2 PE |
Tonbo Biosciences |
Clone RM2–5; RRID:AB_2621728 |
| α-mouse CD2 APC |
Biolegend |
Clone RM2–5; RRID:AB_2563090 |
| α-mouse CD5 Biotin |
Biolegend |
Clone 53–7.3; RRID:AB_312732 |
| α-mouse CD5 FITC |
Biolegend |
Clone 53–7.3; RRID:AB_312734 |
| α-mouse CD5 PE |
Biolegend |
Clone 53–7.3; RRID:AB_312736 |
| α-mouse CD5 APC |
Biolegend |
Clone 53–7.3; RRID:AB_2563929 |
| α-mouse CD3 Biotin |
Biolegend |
Clone 17A2; RRID:AB_2563946 |
| α-mouse CD3 FITC |
Biolegend |
Clone 17A2; RRID:AB_312661 |
| α-mouse CD3 PE |
Tonbo Biosciences |
Clone 17A2; RRID:AB_2621731 |
| α-mouse CD3 APC |
Tonbo Biosciences |
Clone 17A2; RRID:AB_2621538 |
| α-mouse CD8a Biotin |
Tonbo Biosciences |
Clone 53–6.7; RRID:AB_2621638 |
| α-mouse CD8a FITC |
Tonbo Biosciences |
Clone 53–6.7; RRID:AB_2621671 |
| α-mouse CD8a PE |
Tonbo Biosciences |
Clone 53–6.7; RRID:AB_2621741 |
| α-mouse CD8a APC |
Biolegend |
Clone 53–6.7; RRID:AB_312751 |
| α-human/mouse B220 Biotin |
Tonbo Biosciences |
Clone RA3–6B2; RRID:AB_2621644 |
| α-human/mouse B220 FITC |
Tonbo Biosciences |
Clone RA3–6B2; RRID:AB_2621690 |
| α-human/mouse B220 APC |
Tonbo Biosciences |
Clone RA3–6B2; RRID:AB_2621574 |
| α-human/mouse B220 Percp-Cy5.5 |
Tonbo Biosciences |
Clone RA3–6B2; RRID:AB_2621892 |
| α-human/mouse B220 PE |
Tonbo Biosciences |
Clone RA3–6B2; RRID:AB_2621764 |
| α-mouse CD45.1 FITC |
Biolegend |
Clone A20; RRID:AB_313494 |
| α-mouse CD45.1 PE-Cy7 |
Tonbo Biosciences |
Clone A20; RRID:AB_2621850 |
| α-mouse CD45.2 VioletFluor 450 |
Tonbo Biosciences |
Clone 104; RRID:AB_2621950 |
| α-mouse Ter119 Biotin |
Tonbo Biosciences |
Clone TER-119; RRID:AB_2621651 |
| α-mouse Ter119 FITC |
Tonbo Biosciences |
Clone TER-119; RRID:AB_2621720 |
| α-mouse Ter119 PE |
Tonbo Biosciences |
Clone TER-119; RRID:AB_2621802 |
| α-mouse Ter119 Brilliant Violet 510 |
BD Biosciences |
Clone TER-119; RRID:AB_2738532 |
| α-mouse Ter119 APC |
Tonbo Biosciences |
Clone TER-119; RRID:AB_2621609 |
| α-mouse CD71 FITC |
eBioscience |
Clone R17217; RRID:AB_465124 |
| α-mouse c-kit APC-eFluor780 |
eBioscience |
Clone 2B8; RRID:AB_1272177 |
| α-mouse CD117 MicroBeads |
Miltenyi Biotec |
Clone 3C1; RRID:AB_2753213 |
| α-mouse CD45 MicroBeads |
Miltenyi Biotec |
Clone 30F11; RRID:AB_2877061 |
| Anti-APC magnetic microbeads |
Miltenyi Biotec |
RRID:AB_244367 |
| α-mouse Sca1 Percp-Cy5.5 |
Thermo Fisher Scientific |
Clone D7; RRID:AB_914372 |
| α-mouse CD150 PE |
Biolegend |
Clone TC15–12F12.2; RRID:AB_313682 |
| α-mouse CD150 PE-Cy7 |
Biolegend |
Clone TC15–12F12.2; RRID:AB_439796 |
| α-mouse CD150 APC |
Biolegend |
Clone TC15–12F12.2; RRID:AB_493461 |
| α-mouse CD48 APC |
Biolegend |
Clone HM48–1; RRID:AB_571996 |
| α-mouse CD48 PE-Cy7 |
Biolegend |
Clone HM48–1; RRID:AB_2075049 |
| α-mouse CD48 Alexa Fluor 700 |
Biolegend |
Clone HM48–1; RRID:AB_10612754 |
| α-mouse CD34 Biotin |
Thermo Fisher Scientific |
Clone RAM34; RRID:AB_466426 |
| α-mouse CD43 PE |
BD Biosciences |
Clone S7; RRID:AB_394748 |
| α-mouse IgM APC |
Thermo Fisher Scientific |
Clone Il/41; RRID:AB_469458 |
| α-mouse Mac-1 (CD11b) APC-efluor780 |
Thermo Fisher Scientific |
Clone M1/70; RRID:AB_1603193 |
| α-mouse CD16/32 (FcγRIII/II) BV510 |
Biolegend |
Clone 93; RRID:AB_2563692 |
| α-mouse CD45 FITC |
Tonbo Biosciences |
Clone 30-F11; RRID:AB_2621689 |
| Streptavidin PE-Cy7 |
Fisher Scientific |
BDB557598 |
| α-mouse CD4 APC |
Biolegend |
Clone GK1.5; RRID:AB_312697 |
| α-mouse/human CD44 PE-Cy7 |
Biolegend |
Clone IM7; RRID:AB_830787 |
| α-mouse CD25 PerCP-Cy5.5 |
Tonbo Biosciences |
Clone PC61.5; RRID:AB_2621889 |
| α-GLAST APC |
Miltenyi Biotec |
Clone ACSA-1; RRID:AB_2660783 |
| α-GOT1 |
Abcam |
Clone C1; Cat. ab239487 |
| α-GOT2 |
Proteintech |
Polyclonal; RRID:AB_2247898 |
| α-Histone H3 |
Abcam |
Polyclonal; RRID:AB_302613 |
| α-mouse IgG, HRP-linked |
Cell Signaling Technology |
Polyclonal; RRID:AB_330924 |
| α-rabbit IgG, HRP-linked |
Cell Signaling Technology |
Polyclonal; RRID:AB_2099233 |
|
|
Chemicals, Peptides, and Recombinant Proteins
|
| MethoCult™ GF M3434 |
STEMCELL Technologies |
Cat. M3434 |
| Cyclophosphamide |
Amneal Pharmaceuticals |
N/A |
| G-CSF (Neupogen) |
Amgen |
N/A |
| HI bovine serum |
Thermo Fisher Scientific |
Cat. 26170043 |
| Penicillin-Streptomycin solution |
Fisher Scientific |
Cat. SV30010 |
| Penicillin-Streptomycin-Glutamine (100X) |
Thermo Fisher Scientific |
Cat. 10378016 |
| DMEM, high glucose |
Thermo Fisher Scientific |
Cat. 11965118 |
| DMEM, low glucose |
Thermo Fisher Scientific |
Cat. 11885–092 |
| Silac DMEM Flex Media, no glucose, no phenol red |
Thermo Fisher Scientific |
Cat. A2493901 |
| Ham's F-12 Nutrient Mix |
Thermo Fisher Scientific |
Cat. 11765062 |
| Leibovitz L-15 medium, no phenol red |
Life Technologies |
Cat. 21083–027 |
| Neurobasal medium |
Thermo Fisher Scientific |
Cat. 21103049 |
| Insulin-Transferrin-Selenium-Ethanolamine (ITS -X) (100X) |
Thermo Fisher Scientific |
Cat. 51500056 |
| HEPES (1 M) |
Life Technologies |
Cat. 15630–080 |
| Poly(vinyl alcohol),87–90% hydrolyzed, average mol wt 30,000–70,000 |
Sigma-Aldrich |
Cat. P8136–250G |
| Hank's Balanced Salt Solution (HBSS) |
Thermo Fisher Scientific |
Cat. MT21022CV |
| Recombinant murine thrombopoietin (TPO) |
Peprotech |
Cat. 315–14 |
| Recombinant murine SCF |
Peprotech |
Cat. 250–03 |
| Recombinant human bFGF |
R&D Systems |
Cat. 233-FB-025 |
| Recombinant Human EGF Protein, CF |
R&D Systems |
Cat. 236-EG-200 |
| Recombinant Human IGF-I Protein, CF |
R&D Systems |
Cat. 291-G1–200 |
| BioCoat™ Fibronectin 6-well Clear Flat Bottom TC-Treated Multiwell Plate |
Corning |
Cat. 354402 |
| BioCoat™ Fibronectin 96-well Clear Flat Bottom TC-treated Microplates |
Corning |
Cat. 354409 |
| Corning ultra low attachment surface culture dish, 6-well plate |
Corning |
Cat. 07200601 |
| Acetonitrile, Optima LC/MS Grade |
Fisher Scientific |
Cat. A955–500 |
| Methanol, Optima LC/MS Grade |
Fisher Scientific |
Cat. A456–4 |
| Water, Optima LC/MS Grade |
Fisher Scientific |
Cat. W6–4 |
| L-Aspartic acid (13C4, 99%) |
Cambridge Isotope Laboratories |
Cat. CLM-1801-H-PK |
| L-Aspartic acid-15N |
Sigma-Aldrich |
Cat. 332135–100MG |
| L-Aspartic acid (1,4-13C2, 99%) |
Cambridge Isotope Laboratories |
Cat. CLM-4455-PK |
| L-Asparagine:H2O (15N2, 98%) |
Cambridge Isotope Laboratories |
Cat. NLM-3286–0.5 |
| N-Acetyl-L-aspartic acid-1,2,3,4-13C4
|
Sigma-Aldrich |
Cat. 683647 |
| DL-Glutamic acid (2,3,3,4,4-D5, 97%) |
Cambridge Isotope Laboratories |
Cat. DLM-357-PK |
| L-GLUTAMINE (ALPHA-15N, 98%) |
Cambridge Isotope Laboratories |
Cat. NLM-1016–1 |
| Teriflunomide |
MedChem Express |
Cat. HY-15405 |
| 6-Mercaptopurine Monohydrate |
Toronto Research Chemicals |
Cat. M225450 |
| Tween 80 |
Sigma-Aldrich |
Cat. P1754–1L |
| polyinosinic:polycytidylic acid (pIpC) |
GE Healthcare |
Cat. 27–4732-01 |
| Alt-R® S.p. Cas9 Nuclease V3 |
Integrated DNA Technologies |
Cat. 1081059 |
| 4’,6-diamidino-2-phenylindole |
Sigma-Aldrich |
Cat. D8417–10MG |
| Propidium iodide |
Sigma-Aldrich |
Cat. P4170–25MG |
| Alexa Fluor™ 555 Azide, Triethylammonium Salt |
Thermo Fisher Scientific |
Cat. a20012 |
| O-propargyl-puromycin |
Medchem Source |
Custom synthesis |
| O-propargyl-puromycin |
Thermo Fisher Scientific |
Cat. C10459
|
| Paraformaldehyde |
Fisher Scientific |
Cat. AAJ19943K2 |
| Fetal Bovine Serum |
Gemini Bio-Products |
Cat. 900–208 |
| Saponin |
Sigma-Aldrich |
Cat. 47036–50G-F |
| Urea (powder, BioReagent, for molecular biology) |
Sigma-Aldrich |
Cat. U5378–100G |
| Triton X-100 |
Fisher Scientific |
Cat. BP151–100 |
| DTT |
Sigma-Aldrich |
Cat. 10197777001 |
| NuPAGE LDS Sample buffer (4X) |
Life Technologies |
Cat. NP0007 |
| Membrane, Immun-Blot PVDF |
Biorad |
Cat. 1620177 |
| NuPAGE® MES SDS Running Buffer (20X) |
Life Technologies |
Cat. NP0002 |
| Nupage Transfer Buffer (20x) |
Life Technologies |
Cat. NP00061 |
| SuperSignal West Pico Chemiluminescent Substrate |
Thermo Fisher Scientific |
Cat. 34579 |
| SuperSignal™ West Femto Maximum Sensitivity Substrate |
Thermo Fisher Scientific |
Cat. 34095 |
| Restore™ Western Blot Stripping Buffer |
Thermo Fisher Scientific |
Cat. 21059 |
| Trypsin, Bovine Pancreas |
EMD Millipore |
Cat. 6502–2.5MU |
| Trypsin inhibitor from Glycine max (soybean) |
Sigma-Aldrich |
Cat. T6522–100MG |
| DNase I (grade II, from bovine pancreas) |
Sigma-Aldrich |
Cat. 10104159001 |
| N-2 Supplement (100X) |
Thermo Fisher Scientific |
Cat. 17502048 |
| B-27™ Supplement (50X), serum free |
Thermo Fisher Scientific |
Cat. 17504044 |
| Bovine serum albumin |
Sigma-Aldrich |
Cat. A7030 |
| 2-mercaptoethanol |
Sigma-Aldrich |
Cat. M6250–100ML |
| TRIzol LS Reagent |
Thermo Fisher Scientific |
Cat. 10296–010 |
| Iscript Reverse Transcription Supermix |
Bio-Rad |
Cat. 1708841 |
| iTaq Universal SYBR Green Supermix |
Bio-Rad |
Cat. 172–5124 |
|
|
Critical Commercial Assays
|
| Click-iT® Cell Reaction Buffer Kit |
Life Technologies |
Cat. C10269
|
| LS Columns |
Miltenyi Biotec |
Cat. 130–042-401 |
|
|
Deposited Data
|
| RNAseq analysis |
Signer et al., 2016
|
SRA: PRJNA699097 |
| Microarray analysis |
Krivtsov et al., 2006
|
GEO: GSE3725
|
|
|
Experimental Models: Organisms/Strains
|
|
Rosa26
LSL-Glast
|
this paper |
N/A |
|
Rosa26
LSL-gpASNase1
|
this paper |
N/A |
|
Got1
fl
|
this paper |
N/A |
|
Got2
fl
|
this paper |
N/A |
|
Nat8l
fl
|
Maier et al., 2015
|
N/A |
|
Asnsfl (C57BL/6N-Asnstm1a(EUCOMM)Wtsi/H) |
European Mouse Mutant Archive |
RRID:IMSR_EM: 05307 |
|
Mx1-cre (B6.Cg-Tg(Mx1-cre)1Cgn/J) |
Kühn et al., 1995
|
RRID:IMSR_JAX: 003556 |
|
Vav1-cre (B6.Cg-Commd10Tg(Vav1-icre)A2Kio/J) |
De Boer et al., 2003
|
RRID:IMSR_JAX:008610 |
| Flp (B6;SJL-Tg(ACTFLPe)9205Dym/J) |
Rodriguez et al., 2000 |
RRID:IMSR_JAX:003800 |
|
|
Oligonucleotides
|
| genotyping primers, qPCR primers, sgRNA and donor oligos |
Integrated DNA technologies |
Table S4
|
| Alt-R® CRISPR-Cas9 tracrRNA |
Integrated DNA technologies |
Cat. 1072532 |
|
|
Recombinant DNA
|
| pR26 CAG AsiSI/Mlui cloning vector |
Chu et al., 2016
|
RRID:Addgene_74286 |
| TCM1004 -MGC premier cDNA Clone for Slc1a3
|
Transomics |
Cat. BC066154
|
| Guinea pig Asparaginase 1 coding sequence cloning template |
Integrated DNA technologies |
Table S4
|
|
|
Software and Algorithms
|
| Graphpad Prism v.9.0.1 |
Graphpad |
RRID:SCR_002798 |
| Biorender |
Biorender |
RRID:SCR_018361 |
| ImageJ v1.52a |
Wayne Rasband |
RRID:SCR_003070 |
| Chemdraw v.19.1 |
PerkinElmer |
RRID:SCR_016768 |
| Flowjo |
Flowjo |
RRID:SCR_008520 |
| BD FACSDiva |
BD Biosciences |
RRID:SCR_001456 |
| MultiQuant v.4.1.1 |
SCIEX |
N/A |
| TraceFinder v.4.1 or v.5.0 |
Thermo Scientific |
N/A |
