Figure 5.

Transfection of a PC1 minigene containing the splice-site variant resulted in undetectable full-sized PC1 protein and catalytic activity. (a) Western blot (left and middle panels) and immunoprecipitation analysis (right panel) of HEK-293T cells transfected with empty vector (C) or PC1 minigene containing either WT or splice-site variant (SSV) sequences. Cells used for immunoprecipitation were pulse-labeled for 1 hour. Endo F, endoglycosidase F. (b) Fluorogenic assay of PC1 proteolytic activity using PC1 immunopurified from lysates of αTC1-6 cells transfected as in a. Open circles, empty vector; filled diamonds, PC1 minigene; open squares, PC1 splice-site variant minigene.