The Journal of Neuroscience, March 3, 2004, ():
Different Behavior toward Bovine Spongiform Encephalopathy Infection of Bovine Prion Protein Transgenic Mice with One Extra Repeat Octapeptide Insert Mutation
J. Neurosci. Castilla et al.
24: 2156
Supplementary material
Files in this Data Supplement:
- Supplementary Fig. 1
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Supplementary Figure 1. Diagram showing the 7OR-PrP gene construct. The 6OR-PrP gene, previously cut with the NcoI restriction enzyme to eliminate three OR, was used to insert 4 artificial octarepeats in tandem. These OR were generated by specific oligonucleotides with extremes compatible with NcoI (see Materials and Methods).
- supplementary Fig. 2
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Supplementary Figure 2. Quantification of the PrPres contents of different inocula. Serial dilutions of brain homogenates were analysed by Western blotting using MAb 2A11. BSE1: pool of BSE material (TSE/08/59) derived from the brainstems of 49 BSE infected cattle and supplied by the VLA; BSE2: inoculum derived from one BSE infected brainstem supplied by the VLA; bo7ORTgBSE1: pools from a first passage of BSE1 inoculum in bo7ORTg mice. Monoclonal antibody 2A11 was used at a 1:1500 dilution. Relative molecular mass in kilodaltons.
- supplementary Fig. 3
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Supplementary Figure 3. A: Brain homogenates from six bo6ORTg or bo7ORTg mice were solubilized in extraction buffer. Samples were pre-cleared by centrifugation and the supernatant was ultracentrifuged after treatment with 0, 1 and 3 M GdN HCl. Soluble (S) and insoluble (I) proteins were subsequently analysed by SDS-Page and Western blotting. B: Western blot analysis of brain homogenates from bo6ORTg or bo7ORTg mice inoculated with: BSE1, BSE2 or first passage BSE1 in bo7ORTg (bo7ORTgBSE1) before (p0) or after passage in bo6ORTg or bo7ORTg mice (p1 or p2). Samples were previously digested with proteinase-K (20 µg/ml). Monoclonal antibody 2A11 was used at a 1:1500 dilution. Molecular weight in kilodaltons.
