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. 2019 Oct 9;4(5):e00630-19. doi: 10.1128/mSphere.00630-19

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Copyright © 2019 Holec et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 4 — Binding of PrP^Sc to SCL does not alter strain emergence from a mixture. Results of Western blotting (A and B) and migration analysis (C and D) of PrP^Sc from PMCAsi reaction mixtures seeded with HY and DY bound separately to SCL (A and C) or bound to SCL as a mixture (B and D) are shown. Negative-control brain PMCA reactions did not amplify PrP^Sc (A and B, lanes 1). Positive-control PMCA reaction mixtures seeded with either HY-infected (lane 8) or DY-infected (lane 2) brain homogenate maintained the 21-kDa (lane 15) or 19-kDa (lane 9) strain-specific unglycosylated PrP^Sc migration pattern, respectively, after 5 rounds of amplification. Positive-control PMCAsi reaction mixtures seeded with both unbound HY and DY resulted in the emergence of HY PrP^Sc at round 5 (lanes 3 to 7). Experimental PMCAsi reaction mixtures seeded with both SCL-HY and SCL-DY bound separately (A, lanes 10 to 14) or together (B, lanes 10 to 14) resulted in HY PrP^Sc emerging by round 5. Migration of 19-kDa and 21-kDa molecular weight marker is indicated on the left of the Western blot. This experiment was repeated a minimum of 3 times with similar results.