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

Comparison of ICAM-1-Fc by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

<p>SDS-PAGE gel electrophoresis of 5 µg of ICAM-1-Fc expressed in HEK293 cells, COS-7 cells or in mouse myeloma NS0 (R&D Systems) cells. 5 µl protein marker (M) was loaded onto the gel. Samples were reduced using DTT (+) or non-reduced (−). Arrows indicate ICAM-1-Fc bands.</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-5

<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

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-3

<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>SA, and the unselected control, NF54. The median of FITC fluorescence as measured by flow cytometry is shown for each serum sample. Pools of human sera from exposed females and males, and unexposed Danes (DK-pool) were used as primary antibodies. VAR2CSA expression was confirmed by staining with VAR2CSA DBL5ε-specific rabbit antibodies and irrelevant rabbit immune serum was used as negative control. Samples were processed and run simultaneously at individual time points

Changes in gene mRNA levels during erythrocytic development in two phenotypically distinct parasites-0

<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

α₂M binding in parasites not expressing HB3VAR06.

<p>(A) Binding of α₂M to erythrocytes infected by eight genotypically and/or phenotypically different parasite lines, measured by flow cytometry. Control sample labeling (secondary antibody only) is indicated by gray shading. (B) Rosetting frequencies of erythrocytes infected by six genotypically or phenotypically different parasite lines at increasing concentrations of α₂M but without serum, measured by flow cytometry. Means and SD relative to rosetting in the presence of serum are indicated. (C) <i>Ex vivo</i> binding of α₂M (left) and IgM (right) to erythrocytes infected by a <i>P</i>. <i>falciparum</i> patient isolate (P25). (D) Correlation of <i>ex vivo</i> binding of α₂M and IgM to erythrocytes infected by <i>P</i>. <i>falciparum</i> parasites from 12 patients with uncomplicated malaria. Isolate P25 shown in panel C is indicated by an arrow.</p