KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal Rheb antibody | Lab made | N/A |
| Rabbit monoclonal pS6K (Thr389) antibody | Cell Signaling Technology | Cat#9234 |
| Rabbit monoclonal S6K antibody | Cell Signaling Technology | Cat#2708 |
| Rabbit polyclonal pS6 (Ser240/244) antibody | Cell Signaling Technology | Cat#2215 |
| Rabbit monoclonal S6 antibody | Cell Signaling Technology | Cat#2217 |
| Rabbit polyclonal p4EBP1 (Thr37/46) antibody | Cell Signaling Technology | Cat#9459 |
| Rabbit polyclonal 4EBP1 antibody | Cell Signaling Technology | Cat#9452 |
| Mouse monoclonal Beta actin antibody | Boster Biological Technology | Cat#BM0627 |
| Rabbit polyclonal mTOR antibody | Upstate | Cat#07–231 |
| Rabbit polyclonal pPDH-E1α (Ser293) antibody | Abcam | Cat#ab92696 |
| Rabbit polyclonal pPDH-E1α (Ser300) antibody | Millipore | Cat#AP1064 |
| Rabbit polyclonal pPDH-E1α (Ser232) antibody | Millipore | Cat#AP1063 |
| Mouse monoclonal PDH-E1α antibody | Abcam | Cat#ab110334 |
| Rabbit monoclonal Raptor antibody | Cell Signaling Technology | Cat#2280 |
| Rabbit polyclonal PDP1 antibody | Abcam | Cat#ab59706 |
| Rabbit polyclonal PDP2 antibody | Proteintech | Cat#13404–1-AP |
| Mouse monoclonal c-Myc antibody | Merck | Cat#OP10L |
| Rabbit monoclonal c-Myc antibody | Cell Signaling Technology | Cat#2278 |
| Rabbit His-Tag antibody | Cell Signaling Technology | Cat#2365 |
| Mouse HA-Tag antibody | Cell Signaling Technology | Cat#2367 |
| Rabbit monoclonal MnSOD antibody | Epitomics | Cat#2299–1 |
| Mouse monoclonal GAPDH antibody | Millipore | Cat#MAB374 |
| Mouse monoclonal Beta tubulin antibody | Millipore | Cat#05–661 |
| Mouse monoclonal ARC antibody | Lab made | N/A |
| Rabbit polyclonal Tom20 antibody | Santa Cruz Biotechnology | Cat#sc-136211 |
| Rabbit polyclonal Tom40 antibody | Proteintech | Cat#18409–1-AP |
| Rabbit polyclonal pAKT (Ser473) antibody | Cell Signaling Technology | Cat#9271 |
| Rabbit polyclonal AKT antibody | Cell Signaling Technology | Cat#9272 |
| Rabbit monoclonal TSC2 antibody | Cell Signaling Technology | Cat#3990 |
| Rabbit monoclonal pAMPKα (Thr172) antibody | Cell Signaling Technology | Cat#2535 |
| Rabbit polyclonal AMPKα antibody | Cell Signaling Technology | Cat#2532 |
|
Chemicals, peptides, and recombinant proteins | ||
| Rapamycin | Sigma-Aldrich | Cat#PHZ1233 |
| Torin1 | Tocris Bioscience | Cat#4247 |
| KCl | Sigma-Aldrich | Cat#P5405 |
| Bicuculline | Tocris Bioscience | Cat#0130 |
| L-Lactate | Sigma-Aldrich | Cat#L7022 |
| Recombinant Human/Murine/Rat BDNF | PeproTech | Cat#450–02 |
| GTPγS | Millipore | Cat#20–176 |
| GDP | Millipore | Cat#20–177 |
| ATP Assay Kit | Biovision, Inc, CA, USA | Cat#791–100 |
| Acetyl-CoA Assay Kit | Biovision, Inc, CA, USA | Cat#K317–100 |
| Mitochondria Isolation Kit | Beyotime, China | Cat#C3601/3606 |
| PDH Enzyme activity kit | Abcam | Cat#ab109902 |
| Serine/Threonine Phosphatase Assay kit | Promega | Cat#V2460 |
| Duolink In Situ Red Starter Kit Mouse/Rabbit | Sigma-Aldrich | Cat#DUO92101 |
| Pyruvate Assay Kit | Biovision, Inc, CA, USA | Cat#K609–100 |
| Lactate Assay Kit | Biovision, Inc, CA, USA | Cat#K607–100 |
| ADP Assay Kit | Abcam | Cat#ab83359 |
| AMP Assay Kit | Abcam | Cat#ab273275 |
| Creatine Assay Kit | Abcam | Cat# ab65339 |
| ELISA Kit for phospho-creatine | Cloud Clone | Cat#CEV808Ge |
|
Experimental models: organisms/strains | ||
| Floxed Rheb | Zou et al., 2011 | N/A |
| Rosa26-Rheb(S16H) | Zou et al., 2011 | N/A |
| Rosa26-Rheb(Wildtype) | This paper | N/A |
| Floxed mTOR | The Jackson Laboratory | JAX: 011009 |
| Floxed Raptor | The Jackson Laboratory | JAX: 013191 |
| Albumin-cre | The Jackson Laboratory | JAX: 003574 |
| Nestin-cre | Tronche, et al., 1999 | N/A |
| CaMKII-cre | Casanova, et al., 2001 | N/A |
|
Oligonucleotides | ||
| Primer: Pgc1α real-time PCR Forward: CTCCCTGTGGATGAAGACGG |
This paper | N/A |
| Primer: Pgc1α real-time PCR primer Reverse: ATCCTGTGATTGTGATTTGC |
This paper | N/A |
| Primer: Ndufs8 real-time PCR primer Forward: TGGCGGCAACGTACAAGTAT |
This paper | N/A |
| Primer: Ndufs8 real-time PCR primer Reverse: CCTCGGATGAGTTCTGTCCA |
This paper | N/A |
| Primer: Cox5 real-time PCR primer Forward: GGGTCACACGAGACAGATGA |
This paper | N/A |
| Primer: Cox5 real-time PCR primer Reverse: GGAACCAGATCATAGCCAACA |
This paper | N/A |
| Primer: Atp5g1 real-time PCR primer Forward: AGTTGGTGTGGCTGGATCA |
This paper | N/A |
| Primer: Atp5g1 real-time PCR primer Reverse: GCTGCTTGAGAGATGGGTTC |
This paper | N/A |
| Primer: Rheb real-time PCR primer Forward: AAGTCCCGGAAGATCGCCA |
This paper | N/A |
| Primer: Rheb real-time PCR primer Reverse: GGTTGGATCGTAGGAATCAACAA |
This paper | N/A |
| Primer: Arc real-time PCR primer Forward: AAGTGCCGAGCTGAGATGC |
This paper | N/A |
| Primer: Arc real-time PCR primer Reverse: CGACCTGTGCAACCCTTTC |
This paper | N/A |
| siRNA targeting sequence: TSC2 siRNA 1# sense: GCACCUCUACAGGAACUUUTT |
This paper | N/A |
| siRNA targeting sequence: TSC2 siRNA 1# anti-sense: AAAGUUCCUGUAGAGGUGCGG |
This paper | N/A |
| siRNA targeting sequence: TSC2 siRNA 2# sense: GCCUCACAGGUGCAUCAUATT |
This paper | N/A |
| siRNA targeting sequence: TSC2 siRNA 2# anti-sense: UAUGAUGCACCUGUGAGGCCA |
This paper | N/A |
| siRNA targeting sequence: Tom20 siRNA sense: GGUCUUACAGCAAACUCUUTT |
This paper | N/A |
| siRNA targeting sequence: Tom20 siRNA anti-sense: AAGAGUUUGCUGUAAGACCTG |
This paper | N/A |
|
Software and algorithms | ||
| ImageJ | Schneider, et al., 2012 | imagej.nih.gov/ij |
| GraphPad Prism ver 6.0 | GraphPad software Inc | www.graphpad.com |