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Cryo-EM sample preparation and data collection Sucrose gradient purified fractions of the PV3-SC8 or wt PV2 VLPs expressed in yeast and mammalian cells, respectively, were pooled, buffer exchanged into DPBS-EDTA using Zeba Spin Desalting Columns (Thermo Fisher Scientific) with a 7 K molecular weight cut-off (MWCO) and concentrated using Amicon Ultra centrifugal filter devices (100 kDa MWCO, Merck Millipore) to a concentration of 0.91.5 mg/ml