Western Blot Analysis of PrP^res in the Brains of Transgenic Mice Infected with H-Type or BSE-Type Agents

<div><p>(A) Primary or secondary (lane 6) transmission to tgBov mice. BSE-type inocula include cattle BSE no. 3 (lanes 3 and 7), sheep BSE ARQ no. 1 and ARR no. 1 (lanes 8 and 9), goat BSE (lane 10), and vCJD (lane 11). The PrP^res profiles of both H-type and BSE agents in cattle (lanes 2 and 3) are essentially similar to those in tgBov mice (lanes 4–11). Brain tissue equivalent loaded: 2.5 mg in lane 2; 0.15 mg in lane 3; 0.5 mg in lanes 4–12. MI, mock-infected brain; MM, molecular markers.</p><p>(B) Primary transmission to tgOv mice. H-type agent shows a distinct PrP^res pattern in the brain (Br) compared to BSE agents (lane 9, BSE no. 3; lane 10, goat BSE; lane 11, vCJD). Note the lack of PrP^res signal in the spleen (Sp) of H-type–infected mice (lane 7), unlike that in BSE-infected mice (lane 8). Brain or spleen tissue equivalent loaded: 3 mg in lane 2; 0.15 mg in lane 3; 0.5 mg in lanes 4–6; 2 mg in lanes 7–12.</p><p>(C and D) Ratio of high- and low-molecular-mass PrP^res glycoforms in the brains of tgBov (C) and tgOv (D) mice following challenge with H-type or BSE agents (data plotted as means ± SEM). H-type isolates are represented as triangles (no. 1, blue; no. 2, orange; no. 3, pink; and no. 5, black) and BSE agents as squares (BSE no. 3, red; sheep BSE ARQ no. 1, grey; sheep BSE ARR no. 1, yellow; goat BSE, brown; and vCJD, light blue). Secondary transmissions are represented by unfilled symbols of the same colour. Note the strikingly distinct glycoform ratio between H-type and BSE groups in both mouse lines, as reported in cattle [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0020112#ppat-0020112-b009" target="_blank">9</a>].</p>