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Figure 2


Fig. 2. MIC8 is not required for trafficking of MIC3 to the micronemes. (A) Immunofluorescence analysis of MIC8KOi grown for 48 hours in the presence (+) or absence (–) of ATc prior to fixation. Staining was performed with alpha-MIC8, or with alpha-MIC2 as a control (same exposure time for all micrographs). (B) Immunoblot analysis of MIC8KOi and RH parasites grown in the presence or absence of ATc for 48 hours and probed with the indicated antibodies. No significant difference in the expression levels of MIC2, TUB1 or MIC3 could be detected in MIC8KOi cells under the two conditions. (C) Localisation of MIC3 in MIC8KOi parasites grown in the presence of ATc for 48 hours. Parasites were stained with the indicated antibodies. Merged images are shown on the right. Blue stain indicates DAPI (nuclei). (D) Co-localisation studies in the parasite strain T7S4-5, which overexpresses MIC8Ty (see also Fig. 1B). Upper panels: co-localisation study of MIC8Ty and GRASP-RFP (Pfluger et al., 2005) in the parasite strain T7S4-5 transiently transfected with GRASP-RFP demonstrates accumulation of MIC8 in a post-Golgi compartment. Middle panels: co-localisation of MIC8Ty and the propeptide of M2AP (Harper et al., 2006) in early endosomes. Lower panels: co-localisation study of MIC8Ty and MIC3 shows that MIC3 is not localised in early endosomes because of overexpression of MIC8Ty. Right: enlargement of boxed parasite(s) in the merged pictures. Scale bars, 10 µm.





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