Maintenance of P. falciparum FCR3 Expressing IT4VAR20

Plasmodium falciparum laboratory strain FCR3 expressing the IT4VAR20 PfEMP1 (6 (link)) were maintained in blood type O+ with 0.125 μg/ml Albumax II (Invitrogen), 25 mM sodium bicarbonate (Lonza), 2 mM L-glutamine (Sigma-Aldrich), and 0.125 μg/ml gentamycin (Lonza) supplemented RPMI-1640 HEPES medium. The parasites were maintained in a mixture of 5% CO₂ and 2% O₂. Routine antibody and protein selections i were done to ensure proper parasite phenotype and QPCR was used to assess var gene transcription as previously described (6 (link)).

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Petersen J.E., Bouwens E.A., Tamayo I., Turner L., Wang C.W., Stins M., Theander T.G., Hermida J., Mosnier L.O, & Lavstsen T. (2015). Protein C system defects inflicted by the malaria parasite protein PfEMP1 can be overcome by a soluble EPCR variant. Thrombosis and haemostasis, 114(5), 1038-1048.

Publication 2015

Albumax II sodium bicarbonate L-glutamine gentamycin

Antibody Gene transcription Gentamycin Glutamine Hepes Ml ii Parasite Phenotype Plasmodium falciparum Protein i Sodium bicarbonate Strain

Corresponding Organization :

Other organizations : Scripps Research Institute, Universidad de Navarra, University of Copenhagen, Johns Hopkins University

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Variable analysis

independent variables

Plasmodium falciparum laboratory strain FCR3 expressing the IT4VAR20 PfEMP1

dependent variables

Parasite phenotype
Var gene transcription

control variables

Blood type O+
0.125 μg/ml Albumax II (Invitrogen)
25 mM sodium bicarbonate (Lonza)
2 mM L-glutamine (Sigma-Aldrich)
0.125 μg/ml gentamycin (Lonza) supplemented RPMI-1640 HEPES medium
5% CO2 and 2% O2 gas mixture

controls

Routine antibody and protein selections to ensure proper parasite phenotype
QPCR to assess var gene transcription

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