Figure 6
Microneme secretion in PPM5C knockout strain. Extracellular parasites of the parental (par), knockout (∆ppm5c) and complemented (∆ppm5c.cp) strains were incubated for 10 minutes with or without ethanol and spun down. Supernatant, which contains secreted antigens, and pellet, which contain whole parasites, were processed for western blots. Secreted microneme protein Mic2 and secreted dense granule protein Gra1 was detected in the supernatant (top two rows). In the absence of ethanol constitutive and baseline secretion is detected, while ethanol induces calcium dependent microneme secretion. To confirm equal number of parasites protein extract from the parasite pellet was probed with the surface antigen Sag1. Additionally, we probed for Mic2 and Gra1 in extracts from the parasite pellets to confirm equal levels of the proteins across strains tested. All original blots are shown in Supplemental Fig. S5.
