Figure 3.

(A) HDex1p expression and aggregation in COS cells. COS-1 cells transfected with HDex1-Q20, -Q27, -Q32, -Q37, -Q39, -Q40, -Q45, -Q51, and -Q93 constructs were harvested 42 h after transfection and lysed, and the crude cell lysates were separated by centrifugation into an insoluble (pellet) and a soluble (supernatant) fraction (see Materials and Methods). Ten micrograms of total protein from each soluble fraction and 10 μl of each insoluble fraction were run on separate SDS gels and immunoblotted with anti-HD1 serum. ← in the Upper gel marks the origin of electrophresis. (B) Electron micrographs of negatively stained immunogold-labeled HDex1p aggregates. (Left) Protein aggregates formed from purified GST-HDex1-Q51 protein after cleavage with factor Xa. (Right) Aggregated HDex1 protein isolated from HDex1-Q51-expressing COS-1 cells. The sizes of the gold particles were 10 and 5 nm, respectively. (Bar = 100 nm.)