KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal anti-ALT2 | Sigma-Aldrich | cat#HPA051514; RRID:AB_2681516 |
| Rabbit monoclonal anti-GPT | Abcam | cat#ab202083 |
| Mouse monoclonal anti-OXPHOS Rodent Cocktail | Abcam | cat#ab110413; RRID:AB_2629281 |
| Mouse monoclonal anti-α-Tubulin | Sigma-Aldrich | cat#A11126; RRID:AB_2534135 |
| Mouse monoclonal anti-GAPDH | Invitrogen | cat#AM4300; RRID:AB_2536381 |
| Rabbit polyclonal anti-pSer473 AKT | Cell Signaling Technology | cat#9271; RRID:AB_329825 |
| Rabbit polyclonal anti-pThr308 AKT | Cell Signaling Technology | cat#9275; RRID:AB_329828 |
| Mouse monoclonal anti-AKT | Cell Signaling Technology | cat#2920; RRID:AB_1147620 |
| IRDye 800CW Goat Anti-Rabbit IgG Secondary Antibody | LI-COR | cat#926-32211; RRID:AB_621843 |
| IRDye 680RD Donkey Anti-Rabbit IgG Secondary Antibody | LI-COR | cat#926-68073; RRID:AB_10954442 |
| IRDye 680RD Goat Anti-Mouse IgG Secondary Antibody | LI-COR | cat#926-68070; RRID:AB_10956588 |
| Bacterial and virus strains | ||
| Mouse GPT2 shRNA silencing Adenovirus | Vector Biolabs | cat# shADV-260703 |
| Control LacZ- GFP Adenovirus | ThermoFisher | cat# K494100 |
| mouse ATF4 Adenovirus | Vector Biolabs | cat# ADV-253208 |
| b-gal/LacZ Adenovirus | Vector Biolabs | cat# 1080 |
| Biological samples | ||
| Human cadaveric donor liver tissue | Midwest Transplant Services | N/A |
| Pre/Post RYGBS liver samples | Bariatric Surgery Program at Barnes-Jewish Hospital (St. Louis, Missouri) | N/A |
| Blood samples | Clinical Research Unit, Washington University in St. Louis School of Medicine | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| DMEM - Dulbecco’s Modified Eagle Medium | ThermoFisher | cat#11960085 |
| Fetal Bovine Serum | Gibco | cat#26140-079 |
| Collagenase from Clostridium histolyticum | Millipore Sigma | cat#C5138 |
| Lipofectamine RNAiMAX | ThermoFisher | cat#13778075 |
| [6,6-2H]glucose | Cambridge Isotope Laboratories Inc. | cat#DLM-349-PK |
| Insulin (Humulin) | Eli Lilly and Company | cat#Humulin R U-100 |
| Glucagon | Millipore Sigma | cat#G2044 |
| RNA-Bee | Tel-Test | cat#CS-501B |
| Tunicamycin | Millipore Sigma | cat#T7765 |
| Penicillin-Streptomycin (10,000 U/mL) | Gibco | cat#15140122 |
| Amphotericin B | Gibco | cat#15290026 |
| L-Alanine | Millipore Sigma | cat#A7627 |
| L-Glutamine | Millipore Sigma | cat#G3126 |
| Sodium Pyruvate | Millipore Sigma | cat#P4562 |
| UK5099 | Millipore Sigma | cat#PZ0160 |
| Critical commercial assays | ||
| Glucose and Lactate Analyzer | Yellow Spring Instruments Co. | cat#YSI2300 or cat# YSI2900 analyzer |
| Human Insulin-Specific RIA Kit | Linco Research, Inc. | cat#EZHIASF-14K |
| One-Touch Ultra glucometer | LifeScan | N/A |
| Reverse transcription kit | Thermo Fisher (Applied Biosystems) | cat#4368814 |
| Power SYBR Green | Thermo Fisher (Applied Biosystems) | cat#4367659 |
| ALT (SGPT) Kinetic | Teco Diagnostics | cat#A524-150 |
| Erenna Immunoassay System | Singulex | N/A |
| Glucose Oxidase Activity Assay Kit | Millipore Sigma | cat#GAGO20 |
| Thermo Scientific Vanquish Horizon UHPLC | Thermo Scientific | cat#IQLAAAGABHFAPUMZZZ |
| Infinity Triglycerides | Thermo Scientific | cat#TR22421 |
| Infinity Cholesterol | Thermo Scientific | cat#TR13421 |
| Deposited data | ||
| RNA sequencing data | This paper | GSE199975 |
| Experimental models: Cell lines | ||
| Huh7 Cells | Glow Biologics | cat# GBTC-099H |
| Experimental models: Organisms/strains | ||
| Mouse - diet-induced obese mice in the C57BL/6J strain | Jackson Laboratories | Stock# 380050 |
| Mouse – lean controls for diet-induced obese mice in the C57BL/6J strain | Jackson Laboratories | Stock# 380056 |
| Mouse Littermate db/db and heterozygous (db/+) | Jackson Laboratories | Stock# 000697 |
| Mouse Gpt2tm1e(KOMP)Wtsi –“Gpt2 knockout” | Knockout Mouse Project Repository | project ID CSD24977 |
| Mouse B6.Cg-Speer6-ps1Tg(Alb-cre)21Mgn/J –“Alb-Cre” | Jackson Laboratories | Stock# 003574 |
| Mouse B6.Cg-Tg(ACTFLPe)9205Dym/J -“Actin-Flp” | Jackson Laboratories | Stock# 005703 |
| Mouse Gpt2 floxed - generated by crossing Gpt2 knockout and Actin-Flp | This paper | N/A |
| Mouse LS-Gpt2−/− generated by crossing Gpt2 floxed with Alb-Cre | This paper | N/A |
| Oligonucleotides | ||
| ATF4 ASO | Ionis Pharmaceuticals (Carlsbad, CA). | ID: 489707 |
| Scrambled ASO | Ionis Pharmaceuticals (Carlsbad, CA). | ID: 141923 |
| Human GPT2 siRNA | Thermo Fisher | Cat# AM16708; ID# 112332 |
| Human negative control siRNA | Thermo Fisher | Cat# 4404021 |
| PCR Oligonucleotides can be found in Table S3 | N/A | N/A |
| Software and algorithms | ||
| Prism Version 9 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Image Studio Lite Ver 5.2 | Licor | https://www.licor.com/bio/image-studio-lite/ |