Localisation and expression of APC3-GFP in mitotic cells. (A) Live imaging or (B to E) fixed IFA of APC3-GFP with centrin, α-tubulin (α-Tub) or γ-tubulin (γ-Tub) antibodies in schizonts, microgametocytes (gametocytes) and gametes. DIC: Differential interference contrast, Merge: GFP and Hoechst. Scale bar = 5 μm. Live cell imaging revealed cytoplasmic localisation of APC3-GFP in schizonts and microgametocytes (A). APC3-GFP was found to localise to specific foci in the cytoplasm. Co-staining with antibodies against the known centrosomal markers, centrin and ɣ-tubulin, revealed that in schizonts a single focus of APC3-GFP was present associated with each nucleus, which partially co-localised with a single focus of centrin and γ-tubulin; this is highly persuasive of an association of APC3 with the CPs in this stage of the life cycle (B). During male gametogenesis, multiple MTOC-like structures were observed with centrin and γ-tubulin antibodies. A few of these exhibited partial colocalisation with APC3-GFP foci but the majority did not ( C and D).

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