Fig. 6.

Validation of calpain-dependent cytoskeletal processing during parasite rupture. A, Western blot analysis from uninfected RBC (U), iRBC throughout the life cycle (10–40 hpi), and postrupture vesicle (R) membranes was used to validate the proteolytic processing of candidate proteins. Analysis of α-spectrin, β-spectrin, and ankyrin-1 showed major degradation of target proteins only in the postrupture vesicle proteome sample whereas many proteins, such as flotillin (shown as loading controls underneath each blot) showed no proteolytic processing. B, RBC membranes treated for 30 min with activated human calpain-1 were able to recapitulate the processing observed in postrupture vesicles. Western blot analysis of calpain-mediated processing is shown for α-spectrin, β-spectrin, and ankyrin-1.