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HeLa PINK1-GFP knock-in cell line
onA HeLa cell line expressing PINK1 fused to GFP from the endogenous locus was generated using CRISPR–Cas9 nickase–mediated genome editing with homology-directed repair. GFP-positive clones were isolated by fluorescence-activated cell sorting.
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Biochemical analysis of PINK1 activation and ubiquitination signaling upon GTPP treatment
onUbiquitinated proteins were enriched using Halo-m-DSK ubiquitin capture, and GFP-PINK1 complexes were isolated by IP, followed by SDS–PAGE and immunoblotting to assess PINK1 activation and downstream ubiquitination signaling upon GTPP treatment
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Tabular data from Mitochondrial Proteostasis Cryo-ET Analysis
onTabular output data from the Mitochondrial Proteostasis Cryo-ET Analysis as demonstrated in Ehses et al., 2026.
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Molecular dynamics flexible fitting output of mHsp60 models into in situ density maps
onMDFF simulations were performed to evaluate how well atomic models of mHsp60 single rings in different nucleotide states fit the in situ subtomogram averaging density maps by RMSD-based assessment of model–map agreement.
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Fluorescence microscopy analysis of PINK1 localization upon GTPP treatment
onPINK1-GFP Knock-in cells were stained with the mitochondrial membrane potential–dependent dye MitoSpy Red CMXRos and nuclear DNA dye Hoechst 33342 following treatment with 10µM G-TPP over multiple time points.
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ADP-bound human mtHsp60:mtHsp10 single-ring complex (C7), SPA reconstruction
onCryo-EM SPA density-map of a ADP-bound human mtHsp60:mtHsp10 single-ring complex, SPA reconstruction, from recombinant human mHsp60 and mHsp10 complexes, reconstituted in vitro in the presence of ATP.
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ATP-bound human mtHsp60:mtHsp10 double-ring complex (C7), SPA reconstruction
onCryo-EM SPA density-map of a ATP-bound human mtHsp60:mtHsp10 double-ring complex (C7), SPA reconstruction, from recombinant human mHsp60 and mHsp10 complexes, reconstituted in vitro in the presence of ATP.
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ATP-bound human mtHsp60:mtHsp10 single-ring complex (C7), SPA reconstruction
onCryo-EM SPA density-map of a ATP-bound human mtHsp60:mtHsp10 single-ring complex, SPA reconstruction, from recombinant human mHsp60 and mHsp10 complexes, reconstituted in vitro in the presence of ATP.
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ATP-bound human mtHsp60:mtHsp10 half-football complex (C7), SPA reconstruction
onCryo-EM SPA density-map of a ATP-bound human mtHsp60:mtHsp10 half-football complex, from recombinant human mHsp60 and mHsp10 complexes, reconstituted in vitro in the presence of ATP.
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ATP-bound human mtHsp60:mtHsp10 bullet complex (C7), SPA reconstruction
onCryo-EM SPA density-map of a ATP-bound human mtHsp60:mtHsp10 bullet complex, from recombinant human mHsp60 and mHsp10 complexes, reconstituted in vitro in the presence of ATP.
