Upper panel: IF microscopy of fixed PfClpPapt; PfClpPDEAD parasites stained with antibodies against V5 (endogenous PfClpPapt, red), Ty (PfClpPDEAD, green), and DAPI (DNA, blue). The two proteins colocalize in the apicoplast. The z-stack images were deconvolved and projected as a combined single image. Representative image from one of two biological replicates is shown. (Scale bar, 5 μm). Middle panel: IF microscopy of fixed PfClpCwt and PfClpCF1103W parasites stained with antibodies against V5 (PfClpP, red), Ty (PfClpC, green), and DAPI (DNA). The two proteins colocalize in the apicoplast in PfClpCwt parasites but appear in vesicle-like structures in PfClpCF1103W parasites. The z-stack images were deconvolved and projected as a combined single image. Representative images from one of two biological replicates are shown. (Scale bar, 5 μm.) Lower panel: IF microscopy of fixed PfClpSapt parasites that were incubated for 8 d with aTc (Upper) or without aTc and supplemented with IPP (Lower). Parasites were stained with antibodies against V5 (PfClpS, red), ACP (green), and DAPI (DNA). PfClpS localizes to the apicoplast when aTc is present. Upon aTc removal, PfClpS disappears and ACP appears in vesicle-like structures. The z-stack images were deconvolved and projected as a combined single image. Representative images from one of two biological replicates are shown. (Scale bar, 5 μm.).

Florentin A, Stephens DR, Brooks CF, Baptista RP, Muralidharan V. Plastid biogenesis in malaria parasites requires the interactions and catalytic activity of the Clp proteolytic system. Proc Natl Acad Sci U S A. 2020 Jun 1:201919501.