Figure 3. Two-dimensional gel electrophoresis and blotting of wild-type and mutant PrP captured by g5p and treated with PK or with PK plus PNGase F. A–D: Cell lysates containing PrPT183A (A and C) or PrPF198S (B and D) were treated with PK at 25 μg/ml prior to Western blotting. E-J: Cell lysates containing PrPWt (E and H), PrPT183A (F and I), or PrPF198S (G and J) were treated with PK plus PNGase F prior to Western blotting. PrP spots are highlighted by arrows. The dashed-lines are aligned with molecular weight makers. Blots A, B, E, F, and G were probed with 3F4, whereas blots C, D, H, I, and J were probed with 1E4.