KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| anti-PDHA1 (phospho S293) | Abcam | ab92696 |
| anti-PDK1 | Cell Signaling Technologies | 3062S |
| anti-FLAG | Sigma | F1804 |
| anti-Vinculin | Sigma | V9131 |
| APC-conjugated anti-mouse CD69 | BioLegend | 10909 |
| eFlour450-conjugated anti-CD3e | Thermo Fisher | 48-0032-80 |
| anti-mouse CD3e | BD Pharmingen | 553057 |
| anti-mouse CD28 | BD Pharmingen | 553294 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| AZD7545 | Selleck Chemicals | S7517 |
| Sodium pyruvate | Sigma | P2256 |
| Sodium L-lactate | Sigma | L7022 |
| Duroquinone | Sigma | D223204 |
| Metformin hydrochloride | Sigma | PHR1084 |
| FCCP (carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone) | Sigma | C2920 |
| Rotenone | Sigma | R8875 |
| Oligomycin A | Sigma | 75351 |
| Antimycin A | Sigma | A8674 |
| Gramicidin from Bacillus aneurinolyticus (Bacillus brevis) | Sigma | G5002 |
| TMRE (tetramethylrhodamine, ethyl ester) | Thermo Fisher | T669 |
| Critical Commercial Assays | ||
| NAD/NADH-Glo Assay kit | Promega | G9072 |
| ADP/ATP Ratio Assay Kit | Sigma | MAK135 |
| TMRE (tetramethylrhodamine, ethyl ester) assay kit | Abcam | ab113852 |
| Ethanol Assay Kit | Sigma | MAK076 |
| Dynabeads Untouched Human CD4 T Cells kit | Thermo Fisher | 11346D |
| Pan-T cell isolation kit | Miltenyl Biotec | 130-095-130 |
| Deposited Data | ||
| Raw image data | This manuscript | http://dx.doi.org/10.17632/6zpghspxgs.1. |
| Experimental Models: Cell Lines | ||
| Human: 143B | ATCC | CRL-8303 |
| Human: A172 | ATCC | CRL-1620 |
| Human: A549 | ATCC | CCL-185 |
| Mouse: C2C12 | ATCC | CRL-1772 |
| Human: H1299 | ATCC | CRL-5803 |
| Human: HeLa | ATCC | CCL-2 |
| Human: MDA-MB-231 | ATCC | HTB-26 |
| Mouse: PSC | This manuscript | N/A |
| Human: Primary CD4+ T cells | This manuscript | N/A |
| Mouse: Primary T cells | This manuscript | N/A |
| Experimental Models: Organisms/Strains | ||
| Mouse: NU/NU Nude Mouse (Crl:NU-Foxn1nu) | Charles River | 088 |
| S. cerevisiae: CEN.PK 5D (MATa ura3–52 HIS3, LEU2 TRP1 MAL2–8c SUC2) | This manuscript | N/A |
| Oligonucleotides | ||
| sgRNA sequence targeting PDK1 #1 (5’ GCTCACGTACCACTCGGCAG 3’) | Horlbeck et al., 2016 | hCRISPRi-v2.1 |
| sgRNA sequence targeting PDK1 #2 (5’ GACGTCCCTCACGTACCACT 3’ | Horlbeck et al., 2016 | hCRISPRi-v2.1 |
| sgRNA sequence non-targeting control (5’ GGGAACCACATGGAATTCGA 3’) | Horlbeck et al., 2016 | hCRISPRi-v2.1 |
| Recombinant DNA | ||
| pUC57-LbNOX | Addgene | 75285 |
| pInducer20 | Addgene | 44012 |
| Software and Algorithms | ||
| FlowJo | FlowJo | V10.6.1 |
| Xcalibur™ Software | Thermo Fisher | N/A |
| Prism | GraphPad | 8.2.1 |