Macrophages were treated with VLPs composed of up to 360 monomers of WUPyV recombinant major capsid protein VP1 (MW 40 kDa). VLPs were produced in S. cerevisiae yeast expression system and purified by CsCl density gradient centrifugation as described previously [24 (link), 25 (link)]. The isolated VLPs were dissolved in PBS, dialysed, mixed with glycerol (1:1), filtered through a 0.2 µm pore size RC syringe filter and stored at -20 °C. VLP structure was verified by examination of the purified proteins using Morgagni-268 electron microscope (FEI, Inc., Hillsboro, OR, USA).
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