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. 2014 Dec 12;8(6):393–403. doi: 10.4161/19336896.2014.983753

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Figure 4. — Addition of 12 ug of in vitro transcribed sheep or white-tailed deer PrP mRNA molecules to seeded-PMCA resulted in enhanced PrPSc or PrPCWD amplification and increased sensitivity by at least a 100-fold in the second round PMCA. Panel A1 and A2 represent first round PMCA, without and with sheep PrP mRNA, respectively, of scrapie seeded PMCA using sheep rPrP^C (VLRQ) as substrate. Panel B1 and B2, represent second round PMCA, without and with sheep PrP mRNA, respectively. Panel C1 and C2 show the third round of PMCA as indicated in the figure (negative and positive controls are on a separate gels and are not shown). Panel D1 and D2 show the effect of white-tailed deer PrP mRNA on the efficiency of first and second round CWD-seeded PMCA using white-tailed rPrP^C as substrate. PMCA results (rPrP^sC or rPrP^CWD positivity) are indicated below each panel. Panel E shows the effect of specific (PrP mRNA) RNA on conversion efficiency in seeded PMCA. Rift Valley fever virus nucleoprotein (NP) mRNA is utilized as a non-specific RNA control. Densitometric analysis of the rPrP^SC bands (Panel F). www.landesbioscience.