Tex1 localization in trophozoite and schizont stages with respect to newly described structures called tethers.

<p>Co-localization of Tex1 (red) with MAHRP2 (green) A) in trophozoite stages and B) in schizont stages. Nuclear DNA was stained with DAPI (blue), Transmission image (DIC), Scale bar: 5 µm.</p

Time points of harvest of synchronized 3D7 <i>in vitro</i> culture.

<p>Synchronized <i>P. falciparum</i> 3D7 parasite culture were harvested in 4 hours interval. Time points of harvest 1–14 (column 1), the corresponding age of synchronized parasites at each time point of harvest (in hours post invasion, column 2) and the corresponding parasite stage (column 3).</p

Co-localization of Tex1 with SBP1.

<p>P27-specific polyclonal rabbit sera was used to detect Tex1 (red). Co-localization was performed using SBP1 polyclonal mouse sera (green). Co-localization was performed in ring (A) trophozoite (B) and schizont stage (C) infected RBCs. Nuclear DNA was stained with DAPI (blue), Transmission image (DIC), Scale bar: 5 µm.</p

Co-localization of Tex1 with MAHRP1.

<p>P27-specific polyclonal rabbit sera was used to detect Tex1 (red). Co-localization was performed using MAHRP1 polyclonal mouse sera (green). Co-localization was performed in ring stage (A) trophozoite (B) and schizont stage (C) infected RBC. Nuclear DNA was stained with DAPI (blue), Transmission image (DIC), Scale bar: 5 µm.</p

Co-localization of Tex1 with Rex1.

<p>P27-specific polyclonal mouse sera (in red) was used to detect TEX1. Rex1 polyclonal rabbit sera (in green). (A) Ring stage parasites; (B) trophozoite stages; (C) schizont stages. Scatter plots show the degree of co-localization of the Tex1 with Rex1 signal. Nuclear DNA was stained with DAPI (blue), Transmission image (DIC), Scale bar: 5 µm.</p

Brefeldin A sensitivity of Tex1 export.

<p>3D7 infected RBC were treated with BFA and fixed (+BFA). Tex1 was stained using P27 or P27A-specific mouse antibodies (in red, upper panel: early trophozoite, middle panel: trophozoite). Tex1 visible inside the parasite in close proximity to the nucleus. A control culture (+ETOH) was incubated with equivalent concentration of ethanol, the solvent of Brefeldin A. In the control culture Tex1 was correctly exported and associated to MC (in red). The nucleus was stained with DAPI (in blue). Transmission image (DIC). scale bar: 5 µm.</p

Immunofluorescence staining of erythrocytes infected by <i>P. falciparum</i> (ring, trophozoites and schizont stages) using P27-specific polyclonal rabbit sera.

<p>P27-specific polyclonal rabbit sera was used to detect Tex1 (green) A) in late ring stages B) in trophozoite stages C) in schizont stages. Nucleus stained with DAPI (blue), transmission picture of the infected red blood cell (DIC) and merged picture of the two signals or the signals merged with transmission picture (merge), Scale bar: 5 µm.</p

Equinatoxin II assay.

<p>A) 3D7 infected RBC lysed with equinatoxin II. Integrity of MCs is demonstrated by the absence of the SBP1 signal after using SBP1 N-terminus specific polyclonal mouse sera (note: N-terminus of SBP1 faces the lumen of MCs). Tex1 signal on the MC surface was obtained with P27-specific polyclonal rabbit sera (in green). B) 3D7 infected RBC lysed with equinatoxin followed by Triton lysis. MC lumen is now accessible for antibodies as shown by the SBP1 signal (in red). Nuclear DNA stained with DAPI (blue), Transmission image (DIC). Scale bar: 5 µm.</p