Localization of PfSUB3 in the parasite by immuno-fluorescence assay. Immunofluorescent staining of ring, trophozoite and schizont stage parasites with anti-PfSUB3 antibody. Cy3-labeled anti-mouse IgG was used as secondary antibody and nuclei were counterstained with DAPI. Cy3 fluorescence signals were seen in schizont stage parasites. PfSUB3 accumulates in the parasitophorous vacuoleAlam A, Bhatnagar RK, Chauhan VS. Expression and characterization of catalytic domain of Plasmodium falciparum subtilisin-like protease 3. Mol Biochem Parasitol. 2012 183(1):84-9

Fusion protein to PFE0355c accumulates in the parasitophorous vacuole, indicating a functional signal sequence but no functional export motif.Sargeant TJ, Marti M, Caler E, Carlton JM, Simpson K, Speed TP, Cowman AF. Lineage-specific expansion of proteins exported to erythrocytes in malaria parasites. Genome Biol. 2006 7(2):R12; PMID:

Parasites were transfected with the GFP chimeras of the 3 genes. Experimental verification of their sub-cellular localization indicated that REX-3 and Hsp40 were exported into the host cell cytosol while the protease accumulated in the parasitophorous vacuole.Sargeant TJ, Marti M, Caler E, Carlton JM, Simpson K, Speed TP, Cowman AF. Lineage-specific expansion of proteins exported to erythrocytes in malaria parasites. Genome Biol. 2006; 7(2):R12.

Protein expression profile of selected P. falciparum proteases. IFAs using polyclonal mouse antisera directed against 12 of the proteases was used to demonstrate protein expression in the asexual blood stages (R, ring; T, trophozoite; iS, immature schizont; mS, mature schizont) and gametocytes (GC stages III, IV and V) as well as in macrogametes (MaG) at 20 min post-activation (in green). The asexual and the sexual blood sstages were visualized via immunolabelling with polyclonal rabbit antisera against PfMSP1 and Pfs230, respectively (in red). Hoechst 33342 staining was used to highlight the parasite nuclei (in blue). Bar, 5 μm. Results are representative of three to four independent experiments.Weißbach T, Golzmann A, Bennink S, Pradel G, Julius Ngwa C. Transcript and protein expression analysis of proteases in the blood stages of Plasmodium falciparum. Exp Parasitol. 2017 Mar 25. [Epub ahead of print]