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2 protocols using got2 sirna

1

Investigating Glutamate Metabolism in Neuroinflammation

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Lysozyme (Sigma #L6394), LPS (Sigma #L2630), Glutamate assay kit (Sigma #MAK004) and Minocycline were purchased from Sigma. TLR4 (Abcam #ab22048), PGP9.5 (Abcam #ab108986), GOT (Abcam #ab221939) and Glutaminase (Abcam #ab156876) antibodies were purchased from Abcam. NFκB p65 (CST #8242), Glutamate dehydrogenase (CST #80063), GOT1 (CST #34423), and GOT2 (CST #71692) antibodies were purchased from Cell Signaling Technology. PEPinh MyD, PEPinh TRIF were purchased from Invivogen. Cu-CPT22 (#4884), TAK-242 (#6587), and Dynasore (#2897) were purchased from Tocris. TRIF siRNA (Thermo, #4457308), GOT2 siRNA (Thermo, #4457308), HMGB1 (Thermo #34-8401-82), IFNα mouse ELISA kit (Thermo #BMS6027) and IL-10 mouse ELISA kit (Thermo #BMS614) were purchased from ThermoFisher Scientific. IFN-β ELISA kit (PBL Assay Science #424001) was purchased from PBL Assay Science and mouse inflammatory cytokines ELISA array kit (Qiagen #336161) was purchased from Qiagen. Cell culture media and reagents were purchased from Thermo (Invitrogen). All other reagents were purchased from Sigma until stated otherwise.
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2

Plasmid and Metabolic Assay Materials

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The pCMV5-Flag-XBP1s plasmid was purchased from Addgene. Human TNF siRNA, GOT2 siRNA and control siRNA were obtained from Thermo Fisher Scientific. The mitochondrial respiration inhibitors Antimycin A, Rotenone and Oligomycin were purchased from Agilent Technologies, Piericidin A was from Cayman Chemical. NAD+ was obtained from Cayman Chemical. α-ketoglutaric acid, succinic acid, L-aspartic acid, α-ketobutyric acid, L-asparagine were from Sigma-Aldrich. Malic acid was from Santa Cruz Biotechnology. MitoTracker Red and MitoTracker Green were purchased from Thermo Fisher Scientific. Assay kits for aspartate, oxaloacetate, NAD+/NADH and phosphatidylcholine were obtained from Sigma-Aldrich. Elisa kit for human TNF was from Thermo Fisher Scientific.
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