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The flow chart of cryo-EM data processing. Starting from a single dataset collected from the mixture of the PI31 and 20S, we segregated the particle images into two major classes – 20S CP with and without the bound PI31, leading to the two final EM maps of 20S CP particle alone at 2.23 Å average resolution and the 20S complexes with PI31 at 2.54 Å average resolution.
