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Cryo-EM grids preparation and images acquisition for GLS2 (K253A) filament Human GLS2 (K253A), 3 M, was mixed with 20 mM glutamine to initiate filament formation

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This was enabled by a dual fluorescence ENL stability reporter (ENL-TagBFP-P2A-mCherry) expressed in KBM7 cells harboring a doxycycline-inducible Cas9 expression cassette (Extended Data Fig

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