Purification of ICAM1-Fc expressed by HEK293 cells.

<p>Data from exp. #1 is shown here as an example of purification of ICAM-1-Fc_HEK293. (<b>A</b>) Dot blot showing 2 µl of cell supernatant at day of harvest, 2 µl diafiltrated supernatant (column input) and 2 µl column run-through. 1.8 µg and two ten-fold dilutions hereof of the eluted ICAM-1-Fc was dotted onto the membrane. ICAM-1-Fc was detected using HRP-conjugated anti-human IgG antibody. (<b>B</b>) Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel electrophoresis of 5 µl protein marker (lane 1), 10 µl column input (lane 2), 10 µl column run-through (lane 3) and 9 µg eluted ICAM-1-Fc (lane 4+5). Samples were reduced using DTT (+) or non-reduced (−). Arrows indicate ICAM-1-Fc bands.</p

Binding of the malaria PfEMP1 antigen DBLβ3_D4 to ICAM-1-Fc.

<p>Concentration-dependent binding of recombinant <i>P. falciparum</i> 3D7 PFD1235w DBLβ3_D4 to ICAM-1-Fc_HEK239, ICAM-1-Fc_COS-7 and ICAM-1-Fc_NS0 (R&D Systems) by ELISA. The binding of DBLβ3_D4 to ICAM-1-Fc_HEK239 was repeated in three independent experiments (mean and standard deviation shown) while the assay using ICAM-1-Fc_COS-7 and ICAM-1-Fc_NS0 (R&D Systems) was done one time each.</p

Reactivity of monoclonal ICAM antibodies.

<p>The reactivity of seven anti-human ICAM-1 specific monoclonal antibodies (clones 15.2, RR1/1, 84H10, LB2, BBIG-I1, 8.4A6 and My13) against ICAM-1-Fc expressed in HEK293, COS-7 or mouse myeloma NS0 (R&D Systems) cells were tested using ELISA. One CD36 specific monoclonal antibody (clone FA6.152) was included as negative control. Data shown are the mean reactivity (three independent experiments) of the antibodies to ICAM-1. Errors indicate S.D.</p

Changes in gene mRNA levels during erythrocytic development in two phenotypically distinct parasites-1

<p><b>Copyright information:</b></p><p>Taken from "Changes in gene mRNA levels during erythrocytic development in two phenotypically distinct parasites"</p><p>http://www.malariajournal.com/content/6/1/78</p><p>Malaria Journal 2007;6():78-78.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1904452.</p><p></p>on of both populations corresponds to the expression of the most abundant transcript, PFL0935c and (PFL0030c), respectively. The most striking difference in transcription is the slightly earlier distinctive peak for (PFL0030c) in NF54VAR2CSA compared to the broader wave-like appearance of transcription in NF54. All measurements at the different time points were normalized against

Changes in gene mRNA levels during erythrocytic development in two phenotypically distinct parasites-4

<p><b>Copyright information:</b></p><p>Taken from "Changes in gene mRNA levels during erythrocytic development in two phenotypically distinct parasites"</p><p>http://www.malariajournal.com/content/6/1/78</p><p>Malaria Journal 2007;6():78-78.</p><p>Published online 12 Jun 2007</p><p>PMCID:PMC1904452.</p><p></p>(A) and (B) and as pie charts in (C) at different time points during the intra-erythrocytic life cycle of NF54 and NF54VAR2CSA. (C) shows only expression of full-length genes. (A) and (B):blue – (was used for monitoring the different stages), green – PFL0935c (dominant transcript in NF54), grey – , red diamonds – (DBL4 primers), red circles – (DBL2 primers), red rectangles – (5' UTR primers). (C): light green – PFL0935c, red – (DBL2 primers). pi – post invasio

The density of knobs on the surface of erythrocytes infected by isolate GH18 after prolonged culture <i>in vitro</i>.

<p>The relationship between time since invasion and IE surface knob density after <i>in vitro</i> culture of isolate GH18 for 8 weeks (A) or 12 weeks (B). All time/density data points <36 h as well as the common regression line (with 95% confidence interval) for the GH18 (note that the <i>ex vivo</i> data points shown in Fig. 2M are included) (C). Individual data points, as well as the linear regression line (with 95% confidence limits) for data points <36 h are shown. Adjusted regression lines (assuming parallelism) after different times of <i>in vitro</i> culture of isolate GH18 (D).</p

The density of knobs on the surface of erythrocytes infected by long-term in vitro isolates of <i>P. falciparum</i> expressing the PfEMP1 protein VAR2CSA.

<p>The relationship between time since invasion (h) and IE surface knob density (µm^–2) on erythrocytes infected by 10 genotypically distinct isolates of <i>P. falciparum</i> (isolate name in brackets), maintained in long-time <i>in vitro</i> culture and selected for expression of the PfEMP1 protein VAR2CSA by regular panning for IE adhesion to CSA (A-J). Individual data points, as well as the linear regression line (with 95% confidence limits) for data points <36 h are shown for each isolate.</p

Identification and characterization of the α₂M-binding domain in HB3VAR06.

<p>(A) Schematic representation of the domain structure of HB3VAR06. Domain nomenclature as described by Rask <i>et al</i>. [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005022#ppat.1005022.ref037" target="_blank">37</a>], as well as the first and last amino acid in each domain are indicated. (B) Binding of α₂M to recombinant HB3VAR06 single-, double-, and triple-domain constructs (labeled as in panel A) as well as to full-length HB3VAR06 (FV6) measured by ELISA. Means and SD are indicated. (C) Inhibition of α₂M binding to HB3VAR06⁺ IEs by anti-sera raised against the N-terminal head structure (D1–D3), DBLξ2 (D8), and full-length HB3VAR06 (FV6), respectively, measured by flow cytometry. Means and SD relative to binding without anti-serum are indicated. (D) Simultaneous labeling of HB3VAR06⁺ IEs by α₂M and IgM (left), measured by flow cytometry. A control experiment with all detecting antibodies present but without α₂M and IgM is shown to the right. (E) Inhibition of IgM binding to HB3VAR06⁺ IEs by increasing concentrations of α₂M, measured by flow cytometry. Means and SD relative to binding in the absence of α₂M are indicated. (F) Inhibition of α₂M binding to HB3VAR06⁺ IEs by increasing concentrations of IgM, measured by flow cytometry. Means and SD relative to binding in the absence of IgM are indicated.</p

Parasite developmental age and infected erythrocyte knob density.

<p>Example atomic force micrographs of erythrocytes infected by a early schizont parasite <i>ex vivo</i> (A) or by a younger VAR2CSA-expressing trophozoite (B). Black scale bars: 1 µm. The inserts show light micrographs of the same infected erythrocytes stained by Giemsa. Close-up micrographs of knobs (C) and of a single knob (D). Black scale bars: 200 nm (C), 75 nm (D).</p