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Methods Plasmids and lentivirus construction To tag the catalytic subunit of human GCL with a mitochondrial-targeting epitope, the 5-end of the full-length (2,023 bp) cDNA-encoding GCL (GenBank accession number NM_012815) was fused with a cDNA fragment encoding the first 32 amino acids of human ornithine transcarbamylase (GenBank accession number NM_000531)

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Anti-mouse or anti-rabbit secondary antibodies (Cell Signaling Technology) HRP-conjugated at 1:5000 dilution were incubated with the membranes for 1 h at room temperature, and visualized using a chemiluminescence system