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Additional file 5: Figure S3. of Iatrogenic Creutzfeldt-Jakob disease with Amyloid-β pathology: an international study

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posted on 08.01.2018, 05:00 authored by Ignazio Cali, Mark Cohen, Stéphane Haїk, Piero Parchi, Giorgio Giaccone, Steven Collins, Diane Kofskey, Han Wang, Catriona McLean, Jean-Philippe Brandel, Nicolas Privat, Véronique Sazdovitch, Charles Duyckaerts, Tetsuyuki Kitamoto, Ermias Belay, Ryan Maddox, Fabrizio Tagliavini, Maurizio Pocchiari, Ellen Leschek, Brian Appleby, Jiri Safar, Lawrence Schonberger, Pierluigi Gambetti
WB profile of resPrPSc from iCJD and sCJD controls using different WB systems for PrP detection. BHs from the frontal cortex (lanes 8-10) and cerebellum (lanes 1-7) treated with 100 U/ml PK (~2000 μg/ml) were probed with indicated Abs. PrP bands were resolved in 15% Tris-HCl, 8.7 cm-long (A-C and E) and 20 cm-long (D) gels, and visualized with near-infrared LI-COR system (A-D and E(i)) or chemiluminescence (E(ii) and E(iii)). A: profiles of resPrPSc from CJD obtained with an 8.7 cm-long gel are similar to those obtained with the 20 cm-long gels in Fig. 2. The unglycosylated resPrPSc ~21 kDa of type 1 (21), ~20 kDa of type i (i), ~19 kDa of type 2 (2) and ~18 kDa (18) are indicated. B: the ~20 kDa of type i (i) and ~21 kDa of type 1 (21) (lanes 3-8), but not the ~19 kDa band of type 2, immunoreact with Ab 12B2. Profile of resPrPSc from iCJDMMi (lane 5) is indistinguishable from that of CJDMVi+2 (lanes 3 and 4), but differs from those of iCJDMM1 and sCJDMM1 (lanes 6-8), as the ~21 kDa band predominates in these three conditions. C: only ~19 kDa (type 2), but not ~21 kDa (type 1) and ~20 kDa (type i) immunoreacted with Tohoku-2 (To-2). An ~18 kDa (18) band in CJDVV2 and CJDMVi+2, and an unidentified lower size fragment (large arrow) in all cases harboring type 2 were also detected. A non-specific band (indicated by three asterisks) was present in all tested samples. D: high resolution gel electrophoresis in 20-cm long gel revealing ~21 kDa (21) and the ~20 kDa band (i and arrowhead). E(i): enhanced image of dashed area in A showing the bands ~21 kDa (21), ~20 kDa (i and arrowhead), and ~19 kDa (19; lane 4). The detection of the resPrPSc ~20 kDa band in addition to the prominent ~21 kDa fragment in CJDMM1 is most likely due to our use of a high resolution electrophoretic systems. E(ii): profiles of unglycosylated resPrPSc from the same cases in E(i) visualized on film by chemiluminescence. Note that the three CJDMM1 (lanes 6-8) show only ~21 kDa bands. E(iii): the ~20 kDa and ~21 kDa bands visible in B (lanes 3-5) resolve as a single band of ~21 kDa (lanes 4 and 5) when visualized on film by chemiluminescence. (PDF 6122 kb)


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