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Metabolic Profiles Point Out Metabolic Pathways Pivotal in Two Glioblastoma (GBM) Cell Lines, U251 and U-87MG
sgRNA constructs and gene expressing codon optimized (OPT) plasmid constructs sgRNA control and constructs against CBS and L2HGDH were designed targeting GTATTACTGATATTGGTGGG (sgControl) (#230080, Addgene), GGAGCAGACAACCTACGAGG (sg CBS -1) (#230081, Addgene), TGGCAGTGCTGGCAGCACGG (sg CBS -2) (#230082, Addgene), GATATAGTCATCGTTGGTGG (sg L2HGDH -1) (#230083, Addgene), and CACCAGACTGGACATAACAG (sg L2HGDH -2) (#230084, Addgene) and constructed in lentiCRISPR v2-Blast (#83480, Addgene) for Blasticidin selection at a final concentration of 5 g/ml
2 / 10 GTX114199 WB Image Various whole cell extracts (30 g) were separated by 10% SDS-PAGE, and the membrane was blotted with Glutathione Reductase antibody (GTX114199) diluted at 1:1000
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Cystathionine therefore appears to be at the nexus of the metabolic response to EGT absence which infers a hitherto unknown but prominent role for cystathionine in directing the cellular metabolic thiol flux (Sheridan et al., 2016)
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