Purified PrP<sup>Sc</sup>, prepared without proteases, causes PrP<sup>C</sup>-dependent spine loss.


<p>(<b>A</b>) Silver stain and Western blot analysis (using anti-PrP antibody D18) of PrP<sup>Sc</sup> purified from scrapie-infected brains without proteases, and mock-purified material from uninfected brains. Lane M, molecular size markers in kDa. Hippocampal neurons from wild-type (WT) mice (<b>B, C</b>) and PrP knockout (<i>Prn-p</i><sup>0/0</sup>) mice (<b>D, E</b>) were treated for 24 hr with 4.4 μg/ml of purified PrP<sup>Sc</sup> (<b>C, E</b>), or with an equivalent amount of material mock-purified from uninfected brains (<b>B, D</b>). Neurons were then fixed and stained with Alexa 488-phalloidin. Scale bar in panel E = 20 μm (applicable to panels B-D). Pooled measurements of spine number (<b>F</b>) and area (<b>G</b>) were collected from 22–25 cells from 4 independent experiments. ***p<0.001 by Student’s t-test; N.S., not significantly different.</p

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