Immunolocalization of PfoTPT to the outside of free, intact apicoplasts. (A) Z-stack of immunofluorescent images (1– 8) showing localization of anti-PfoTPT (red) around apicoplast-targeted GPF (green) in parasites expressing PfACP(leader)-GFP. (B) Electron micrograph of intact apicoplast labeled with anti-PfoTPT antibodies and 20-nm colloidal gold. (C) Tannic acid fixation showing four bounding membranes around isolated apicoplast labeled with anti-PfoTPT antibodies and 10-nm colloidal gold. (Scale bars, 200 nm.)Lowest row: Anti-PfiTPT (red) colocalizes with apicoplast-targeted GPF (green) in parasites expressing PfACP(leader)-GFP.Mullin KA, Lim L, Ralph SA, Spurck TP, Handman E, McFadden GI. Membrane transporters in the relict plastid of malaria parasites. Proc Natl Acad Sci U S A. 2006 103:9572-7. Copyright 2009 National Academy of Sciences, U.S.A.

Indirect immunofluorescence assay on TPT–GFP parasites: TPT–GFP blood stage parasites were stained with primary rabbit anti-ENR polyclonal antibody and visualized by staining with secondary anti-rabbit IgG antibody coupled with Alexa 594. Parasite nuclei are stained with DAPI. In DIC are shown parasitized RBC. Merge shows overlap of TPT–GFP, rabbit anti-enoyl ACP reductase (ENR), DAPI and DIC. Apicoplast area is zoomed to show the colocalization of TPT–GFP with ENR. TPT, which was correctly localized to apicoplast. Plasmodium enoyl ACP reductase (ENR) is well-known apicoplast resident protein.Banerjee T, Jaijyan DK, Surolia N, Singh AP, Surolia A. Apicoplast triose phosphate transporter (TPT) gene knockout is lethal for Plasmodium. Mol Biochem Parasitol. 2012 186(1):44-50. . Copyright Elsevier.