Fig. 4.

The aggregation determinant of CyPrP includes β-sheet 1 and α-helix 1. (a) Diagrams of the of CyPrP^EGFP and several deletion mutants engineered in this study are shown. Numbers indicate residues at the junction of different structural domains (adapted from Zahn et al. 2000). The black box represents the EGFP coding sequence. Aggregation was evaluated by fluorescence microscopy of transiently transfected N2a cells (original magnification ×40). (b) Western blot analysis of the constructs described in (a); extracts containing 100 μg protein from N2a cells were immunostained using antibodies directed against EGFP; mock-transfected cells (lanes 1), CyP-rP^EGFP (lanes 2, expected Mr 54), CyP-rP^EGFPΔOR (lanes 3, expected Mr 49.5), CyPrP^EGFP124stop (lanes 4, expected Mr 38, CyPrP^EGFP124–230 (lanes 5, expected Mr 39), CyPrP^EGFP157stop (lanes 6, expected Mr 42) and CyPrP^EGFP124–157 (lanes 7, expected Mr 33.5). Mr values are indicated on the left. S, supernatant; P, pellet.