The microarray measurements were performed as described in [36 (link)]. Briefly, RNA was extracted using TriZol reagent (Invitrogen, Carlsbad, CA) and the quality and quantity determined by NanoDrop (NanoDrop Technologies). 300 ηg of RNA was used for cDNA synthesis using Sigma WTA2 whole transcriptome amplification kit. 1 μg of cDNA was labeled with Cy3 dye and allowed to hybridize to a custom Agilent array for 17 h followed by washing. The microarray image was taken using a 2 μM scanner and probe intensity values obtained using Agilent Feature Extraction software. Normalization of probe intensities was done using the robust multichip average (RMA) method. The time points were obtained as described above for the RNA-seq measurements and include the time points 6 (n = 3), 24 (n = 3) and 38 (n = 3) hours after time zero.
Ethical approval for the use of human blood in this study was granted by the Institutional Review Boards of the University of South Florida and the University of Notre Dame. All of the blood used for the in vitro culturing of parasites was obtained from healthy adult volunteers and drawn by trained personal from Interstate Blood Bank.
The NF54 strain was originally obtained from the Naval Medical Research Center.
Ethical approval for the use of human blood in this study was granted by the Institutional Review Boards of the University of South Florida and the University of Notre Dame. All of the blood used for the in vitro culturing of parasites was obtained from healthy adult volunteers and drawn by trained personal from Interstate Blood Bank.
The NF54 strain was originally obtained from the Naval Medical Research Center.
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