Subcellular localization of SufS to the apicoplast. Upper panel) Epifluorescent images of live P. falciparum erythrocytic-stage parasites expressing GFP fused to the SufS leader peptide (SufSlp-GFP). The parasites were stained with mitotracker to identify mitochondria and DAPI to identify nuclei. Image z-stacks were deconvolved and then presented as a single combined image. Scale bar = 2 mm. GFP fluorescence localizes to an elongated organelle distinct from the mitochondrion in late ring (top row), late trophozoite or early schizont (middle), and schizont (bottom) stage parasites. Lower panel) Immunofluorescence co-localization of SufSlp-GFP and endogenous ACP. An antibody specific for GFP co-localized with αACP antibodies, demonstrating apicoplast localization in late ring (top panel), late trophozoite or early schizont (middle), and schizont (bottom) stage parasites. The parasites were stained with DAPI to identify nuclei. Image z-stacks were deconvolved and then presented as a single combined image. Scale bar = 2 µm.Gisselberg JE, Dellibovi-Ragheb TA, Matthews KA, Bosch G, Prigge ST. The Suf Iron-Sulfur Cluster Synthesis Pathway Is Required for Apicoplast Maintenance in Malaria Parasites. PLoS Pathog. 2013 9(9):e1003655

Immunofluorescence localisation of PfSufS and PfSufE. (A) Immunofluorescence assay of P. falciparum 3D7 cells using anti-PfSufS Ab with anti-PfHU Ab as apicoplast marker (panel i) or Mitotracker Red (panel ii) indicates targeting of PfSufS to the apicoplast. (B) Co-localisation of PfSufE and apicoplast-targeted GFP observed in the P. falciparum D10 ACPleader-GFP cell line using anti-PfSufE and anti-GFP antibodies (panel i). No overlap of PfSufE signal was observed with Mitotracker Red in P. falciparum 3D7 cells (panel ii) indicating apicoplast localization of PfSufE. PfSufS co-localised with PfHU, an apicoplast DNA condensation protein and no overlapping signals for PfSufS were seen with the mitochondrial marker Mitotracker Red in P. falciparum 3D7 cells (A). PfSufE was localised in the P. falciparum D10-ACPleader-GFP line that targets GFP to the apicoplast. Specific PfSufE signals overlapping with apicoplast-targeted GFP were observed and no overlap of PfSufE signal was seen with Mitotracker Red (B). The localisation of PfSufS and PfSufE exclusively to the apicoplast was thus confirmed.Charan M, Singh N, Kumar B, Srivastava K, Siddiqi MI, Habib S. Sulphur mobilisation for [Fe-S] cluster assembly by the essential SUF pathway in the Plasmodium falciparum apicoplast and its inhibition. Antimicrob Agents Chemother. 2014 Apr 7. [Epub ahead of print]

Immunofluorescence localisation of PfSufS and PfSufE. (A) Immunofluorescence assay of P. falciparum 3D7 cells using anti-PfSufS Ab with anti-PfHU Ab as apicoplast marker (panel i) or Mitotracker Red (panel ii) indicates targeting of PfSufS to the apicoplast. (B) Co-localisation of PfSufE and apicoplast-targeted GFP observed in the P. falciparum D10 ACPleader-GFP cell line using anti-PfSufE and anti-GFP antibodies (panel i). No overlap of PfSufE signal was observed with Mitotracker Red in P. falciparum 3D7 cells (panel ii) indicating apicoplast localization of PfSufE. PfSufS co-localised with PfHU, an apicoplast DNA condensation protein and no overlapping signals for PfSufS were seen with the mitochondrial marker Mitotracker Red in P. falciparum 3D7 cells (A). PfSufE was localised in the P. falciparum D10-ACPleader-GFP line that targets GFP to the apicoplast. Specific PfSufE signals overlapping with apicoplast-targeted GFP were observed and no overlap of PfSufE signal was seen with Mitotracker Red (B). The localisation of PfSufS and PfSufE exclusively to the apicoplast was thus confirmed.Charan M, Singh N, Kumar B, Srivastava K, Siddiqi MI, Habib S. Sulphur mobilisation for [Fe-S] cluster assembly by the essential SUF pathway in the Plasmodium falciparum apicoplast and its inhibition. Antimicrob Agents Chemother. 2014 Apr 7. [Epub ahead of print