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. 2023 Oct 10;10(12):2316–2323. doi: 10.1002/acn3.51919

Figure 1.

Figure 1

Optimization of PMCA for each subtype of sCJD. To evaluate baseline max amplification of each sCJD subtype, serial dilutions of sCJD brain homogenate (BH) were added to 10% w/v BH from transgenic mice overexpressing HuPrPC with either methione or valine at position 129. Three rounds of PMCA were conducted consisting of 144/96/96 PMCA cycles; each PMCA cycle consists of a 20 sec sonication pulse every 29 min and 40 sec of incubation. PMCA amplification was visualized with western blotting after proteinase K treatment (100 μg/μL for 1 h) and probed with the prion protein specific antibody, 6D11 (1:30,000 dilution). The figure shows the results of the third round of PMCA for each sCJD subtype using either 129M or 129V transgenic mice BH as substrate. Normal brain homogenate (NBH) from humanized transgenic mice without PK digestion was loaded as a migration control. Numbers in the left show the position of molecular weight standards.