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. 2012 Jun;86(11):6033–6041. doi: 10.1128/JVI.07027-11

Fig 4.

Fig 4

Characterization after transmission of sCJD(VV2) prions to Tg mice expressing human (Hu) or chimeric (MHu#2) PrP with methionine (M) or valine (V) at polymorphic codon 129. (A) Immunoblot of brain homogenates: lane 1, inoculum; lane 2, Tg(HuPrP,M129)440; lane 3, Tg(MHu#2,M129)22372; lane 4, Tg(HuPrP,V129+/+)152; and lane 5, Tg(MHu#2,V129)6550 mice. Samples were treated with 100 μg/ml of PK for 1 h at 37°C prior to being loaded on gels and probed with the anti-PrP HuM-P antibody. Apparent molecular masses of migrated protein standards are shown in kilodaltons. (B) Proportions of unglycosylated (black), monoglycosylated (gray), and diglycosylated (white) PK-resistant PrP for each construct from multiple samples: 1, sCJD(VV2) (1 sample; 13 replicates); 2, HuPrP(M129) (3 samples; 7 total replicates); 3, MHu#2(M129) (1 sample; 6 replicates); 4, HuPrP(V129) (1 sample; 4 replicates); 5, MHu#2(V129) (5 samples; 14 total replicates). Bars represent means, and error bars represent standard deviations; statistical difference from inoculum (lane 1) indicated above each bar: n.s., not significant; **, P < 0.01. (C to H) Micrographs of brain tissue from ill Tg mice, showing the most dramatic neuropathological changes: the thalamus of Tg(HuPrP,V129)152 mice (C to E) and piriform cortex of Tg(MHu#2,V129)7110 mice (F to H). Samples were stained with H&E (C, F) and stained immunohistochemically for PrPSc (D, G) or glial fibrillary acidic protein (E, H). Bar in panel H represents 100 μm and applies to all micrographs. (I, J) Vacuolation score histogram, an estimate of the brain area occupied by vacuoles, for Tg(HuPrP,V129)152 mice (I) and Tg(MHu#2,V129)7110 mice (J). NC, neocortex; Pir, piriform cortex; Hp, hippocampus; Th, thalamus; Hab, habenula; Hy, hypothalamus; Cd, caudate nucleus; Cb-G, cerebellar cortex, granular cell layer; Bs, brainstem. (K, L) Kaplan-Meier survival graphs for first (black) and second (gray) transmissions of sCJD(VV2) prions in Tg(MHu#2,V129)7110 (second passages of two independent brains shown) (K) and Tg(MHu#2,V129)6550 (L) mice.