Figure 6:

Fiber formation can be templated by yeast cell lysates. A) Cultured yeast cells harboring the desired prion phenotype are grown to mid-log phase and then lysed by bead beating. B) Lysates are diluted into buffer containing low concentrations of purified recombinant denatured protein. C & D) Subsequent rounds of polymerization are seeded by addition of a small volume of the previous reactions, effectively removing cellular material by dilution. E) These lysate templated seeds are used to polymerize a large volume of purified recombinant isotopically enriched protein into the amyloid form. F) This sample serves as a control for the effects of cellular environment on protein structure to complement study of the same protein in cellular lysates.