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. 2003 Apr 8;100(8):4784–4789. doi: 10.1073/pnas.2627989100

Figure 1.

Figure 1

Western blots of brain homogenates from inocula and inoculated Tg5378 and Tg22372 mice. (a) Immunoblots of rPrPSc (Upper) and deglycosylated rPrPSc (Lower) from brain homogenates, inoculated first and second passages, respectively, of sCJD(MM1) (lanes 1–3), fCJD(E200K) (lanes 4–6), FFI (lanes 7–9), and vCJD (lanes 10–12) into Tg5378 mice. In the first lane of each triplet (lanes 1, 4, 7, and 10), PrPSc in homogenates of the diseased human brains is shown; in the second lane of each triplet (lanes 2, 5, 8, and 11), PrPSc in brain homogenates of Tg5378 mice inoculated with diseased human brains is shown; and in the third lane of each triplet (lanes 3, 6, 9, and 12), PrPSc in brain homogenates from Tg5378 mice inoculated with diseased Tg5378 brain homogenates (from first passage of human prions) is shown. (b) Undigested brain homogenates from sCJD (RG) (lane 1); sCJD (EC) (lane 2); sCJD (WP) (lane 3), and Tg22372 mice inoculated with sCJD (RG) (lane 4); sCJD (EC) (lane 5); and sCJD (WP) (lane 6). Lane 7 shows an uninoculated Tg22372 mouse. Lanes 8–14 show the same brain homogenates as lanes 1–7, respectively, but brain homogenates were digested with proteinase K. Lanes 15–17 are the same as lanes 8–10, respectively, except the film was exposed longer to detect the weak band of PrPSc in lane 16. The blot clearly shows the overexpression of PrP in the brains of transgenic mice relative to human brains (compare lane 7 with lanes 1–3). (c) Brain homogenates from sCJD(MM1) (RG) (lanes 1, 5, and 9) and from Tg22372 mice inoculated with sCJD (RG) on first passage (lanes 2, 6, and 10) and second passage (lanes 3, 7, and 11). Brain homogenate from the second passage of sCJD (RG) into Tg5378 mice are shown in lanes 4, 8, and 12. Lanes 1–4 show undigested brain homogenates, lanes 5–8 show brain homogenates after 100 μg/ml PK digestion, and lanes 9–12 show brain homogenates after both PK digestion and PNGase F digestion. The glycoform pattern of PrPSc changes from the original sCJD (RG) pattern (lane 5) to the first passage (lane 6), remains the same on second passage (lane 7) but changes when passaged into Tg5378 mice (lane 8). The gel mobility of deglycosylated rPrPSc does not change (compare lane 9 to lanes 10–12).