PfTRiC is interacting with the PEXEL-positive PfE60 Maurer’s cleft membrane protein in the cytoplasm of 3D7 iRBCs. Fluorescent patterns of iRBCs infected by 3D7/pARL2-PfE60-gfp (live imaging) and of resealed ghosts from 3D7/pARL2-PfE60-gfp iRBCs (GFP fluorescence, in green, and immunodetection using anti-SBP1 antibodies, in red). RBCs and ghost preparations were incubated with DAPI for nucleus labelling. The GFP-tagged PfE60 protein was detected in the membrane of Maurer’s clefts as shown previously for native PfE60.Mbengue A, Vialla E, Berry L, Fall G, Audiger N, Demettre-Verceil E, Boteller D, Braun-Breton C. New export pathway in Plasmodium falciparum-infected erythrocytes: role of the parasite group II chaperonin, PfTRiC. Traffic. 2015 Jan 23. [Epub ahead of print]

PfTRiC is exported to the cytoplasm of P. falciparum iRBCs. (B). Immunofluorescence studies of erythrocyte infected by Pf3D7 at the early (row 1) and mid (row 2) trophozoite stages and at late schizonte stage using PfTRiCθ specific antibodies (in red). DAPI: DAPI staining of the parasite nucleus (in blue). (C) Double IFA labelling of PfTRiCq (in red) and the rhoptry protein PfRhopH2 (in green) in segmented schizonts using specific sera and methanol-acetone fixed infected erythrocytes .Mbengue A, Vialla E, Berry L, Fall G, Audiger N, Demettre-Verceil E, Boteller D, Braun-Breton C. New export pathway in Plasmodium falciparum-infected erythrocytes: role of the parasite group II chaperonin, PfTRiC. Traffic. 2015 Jan 23. [Epub ahead of print]

Validation of rabbit antibodies against PfTRiCθ. (A) Immunofluorescence studies using the rabbit pre-immune (row 1) andPfTRiCθ immune (rows 2 & 3) sera and Pf3D7 parasite culture. PfTRiCθ (PF3D7_0214000). By immufluorescenceno labelling of uninfected or infected erythrocytes was observed using the pre-immune rabbit serum while a labelling specific for infected erythrocytes was observed using the immune serum.Mbengue A, Vialla E, Berry L, Fall G, Audiger N, Demettre-Verceil E, Boteller D, Braun-Breton C. New export pathway in Plasmodium falciparum-infected erythrocytes: role of the parasite group II chaperonin, PfTRiC. Traffic. 2015 Jan 23. [Epub ahead of print]

PfTRiC is exported to the cytoplasm of P. falciparum iRBCs. (B) Immuno-fluorescence studies of erythrocyte infected by Pf3D7 at the early (row 1) and mid (row 2) trophozoite stages and at late schizonte stage using PfTRiCθ specific antibodies (in red). DAPI: DAPI staining of the parasite nucleus (in blue). (C) Double IFA labelling of PPfTRiCθ (in red) and the rhoptry protein PfRhopH2 (in green) in segmented schizonts using specific sera and methanol-acetone fixed infected erythrocytes. PfTRiCθ was detected in the iRBC cytosol from early rings to early schizonts. Labelling of the parasite (and possibly the parasitophorous vacuole) was instead observed for mature stages (mature trophozoites to schizonts). Moreover, following the individualization of merozoites, PfTRiCθ was only detected at the apex of each merozoite. Double labelling immunofluorescence experiments using anti-PfTRiCθ and anti-RhopH2 antibodies.Mbengue A, Vialla E, Berry L, Fall G, Audiger N, Demettre-Verceil E, Boteller D, Braun-Breton C. New Export Pathway in Plasmodium falciparum-Infected Erythrocytes: Role of the Parasite Group II Chaperonin, PfTRiC. Traffic. 201516(5):461-75.

Loss of PfTRiC-θ does not alter export of PfSBP1 or PfREX1. IFA of acetone-fixed ring-stage infected RBC. i) PfTRiC-θ ± 0.5 μM aTc (anhydrotetracycline), ii) HSP101 ± 10 μM trimethoprim (TMP). DAPI stains the parasite nuclei and PfEXP2 delineates the parasite parasitophorous vacuole compartment. Scale bars = 5 μm. Data are representative of two independent assays. DIC = differential interference contrast. Loss of the PfTRiC complex does not alter export of PfSBP1 or PfREX1 (i). This was in contrast to the block in export of PfSBP1 and PfREX1 observed upon TMP removal in the previously characterized PfHSP101-DDD line (ii).Spillman NJ, Beck JR, Ganesan SM, Niles JC, Goldberg DE. The chaperonin TRiC forms an oligomeric complex in the malaria parasite cytosol. Cell Microbiol. 2017 Jan 9.