KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| phospho-S6 (Ser240/244) | Cell Signaling Technology | Cat# 5364; RRID:AB_10694233 |
| S6 | Santa Cruz Biotechnology | Cat# sc-74459; RRID:AB_1129205 |
| phospho-AKT (Ser473) | Cell Signaling Technology | Cat# 4060; RRID:AB_2315049 |
| AKT | Cell Signaling Technology | Cat# 4691; RRID:AB_915783 |
| NPRL2 | Cell Signaling Technology | Cat# 37344; RRID:AB_2799113 |
| mTOR | Cell Signaling Technology | Cat# 2983; RRID:AB_2105622 |
| phospho-mTOR (Ser 2448) | Cell Signaling Technology | Cat# 5536; RRID:AB_10691552 |
| beta-actin | Cell Signaling Technology | Cat# 3700; RRID:AB_2242334 |
| phospho-p70 S6 Kinase (Thr389) | Cell Signaling Technology | Cat# 9234; RRID:AB_2269803 |
| DEPDC5 | Sigma-Aldrich | Cat# HPA055619; RRID:AB_2682864 |
| TSC2 | Santa Cruz Biotechnology | Cat# sc-271314; RRID:AB_10611435 |
| p70 S6 Kinase | Santa Cruz Biotechnology | Cat# sc-8418; RRID:AB_628094 |
| 4E-BP1 | Cell Signaling Technology | Cat# 9644; RRID:AB_2097841 |
| 4E-BP2 | Cell Signaling Technology | Cat# 2845; RRID:AB_10699019 |
| alpha-tubulin | Cell Signaling Technology | Cat# 3873; RRID:AB_1904178 |
| LAMP1 | Enzo Life Sciences | Cat# ADI-VAM-EN001; RRID:AB_10630197 |
| Goat anti-mouse IRDye 680RD | LI-COR Biosciences | Cat# 925–68070; RRID:AB_2651128 |
| Donkey anti-rabbit IRDye 800CW | LI-COR Biosciences | Cat# 926–32213; RRID:AB_621848 |
| Goat anti-mouse Alexa Fluor 488 | Invitrogen | Cat# A32723; RRID:AB_2633275 |
| Donkey anti-mouse Cy5 | Jackson ImmunoResearch | Cat# 715–175-150; RRID:AB_2340819 |
| Donkey anti-rabbit Cy3 | Jackson ImmunoResearch | Cat# 711–165-152; RRID:AB_2307443 |
|
Bacterial and virus strains | ||
| Tsc2-sh-GFP | Di Nardo et al., 2009 | N/A |
| Control sh-GFP for Tsc2 | Flavell et al., 2006 | N/A |
| Depdc5-sh-GFP | Origene | TL508165 |
| Control-sh-GFP | Origene | TR30021 |
|
Chemicals, peptides, and recombinant proteins | ||
| Pentylenetetrazol | Sigma-Aldrich | P6500–25G |
| Poly-D-Lysine Hydrobromide | MP Biomedical | Cat# 102694 |
| Laminin | ThermoFisherScientific | 23017–015 |
| Rapamycin | EMD Millipore | Cat# 53123–88-9 |
| DAPI | ThermoFisherScientific | Cat# 28908 |
| Glycine | Sigma | G8790–100G |
| L-Alanine | Sigma | A7469–25G |
| L-Arginine | Sigma | A8094–25G |
| L-Asparagine monohydrate | Sigma | A7094–25G |
| L-Cysteine | Sigma | C7352–25G |
| L-Glutamine | Sigma | G3126–100G |
| L-Histidine | Sigma | H6034–25G |
| L-Isoleucine | Sigma | I7403–25G |
| L-Leucine | Sigma | L8912–25G |
| L-Lysine Monohydrochloride | Sigma | L8662–25G |
| L-Methionine | Sigma | M5308–25G |
| L-Phenylalanine | Sigma | P5482–25G |
| L-Proline | Sigma | P5607–25G |
| L-Serine | Sigma | S4311–25G |
| L-Threonine | Sigma | T8441–25G |
| L-Tryptophan | Sigma | T8941–25G |
| L-Tyrosine | Sigma | T8566–25G |
| L-Valine | Sigma | VO513–25G |
|
Critical commercial assays | ||
| Beta hydroxybutyrate assay kit | Abcam | ab83390 |
| One touch ultra 2 glucometer | One touch | 024046 |
|
Deposited data | ||
| Metabolomic analysis | This paper; Metabolomics Workbench |
Table S1, Metabolomics Workbench: https://doi.org/10.21228/M88D9X |
|
Experimental models: Cell lines | ||
| HEK293T | ATCC | Cat# CRL-3216, RRID: CVCL_0063 |
| HeLa | Generated in BD Manning lab | Huang et al., 2008 |
|
Experimental models: Organisms/strains | ||
| Sprague-Dawley Rat | Charles River Laboratories | N/A |
| Mouse: Depdc5f/f-SynCre+ | Generated in M Sahin lab | Yuskaitis et al., 2018 |
|
Software and algorithms | ||
| Fiji Software v2.0.0 | Open Source | http://fiji.sc/;RRID:SCR_002285 |
| Image Studio Lite analysis software v5.2.5 | LI-COR | https://www.licor.com/ |
| GraphPad PRISM v9.0 | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
| MetaboAnalyst v5.0. | MetaboAnalyst Software | https://www.metaboanalyst.ca/ |
|
Recombinant DNA | ||
| psPAX2 | Addgene | RRID:Addgene_12260 |
| pMD2.G | Addgene | RRID:Addgene_12259 |