Comparison of wild type and CNBPKO mice infected with Plasmodium chabaudi
Comparison of wild type and CNBPKO mice infected with Plasmodium chabaudi
Wild-type (CNBP vav-iCre -/- ) and  knockout (CNBP vav-iCre +/+) mice were grouped into infected and uninfected cohorts, total four groups. Mice in the infected groups were injected intraperitoneally with 1 x 10^6 Plasmodium chabaudi-infected red blood cells, while uninfected controls received an injection of sterile 1X PBS. Seven days post-infection (7dpi), all mice were euthanized, and spleens were harvested. C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME) were maintained in specific pathogen-free conditions and used in accordance with the Institutional Animal Care and Use Committee.  Intra-peritoneal injection of 1 x10e6 P.chabaudi infected Red Blood Cells. Approximately 15 mg of splenic tissue was homogenized in RLT buffer from the RNeasy Kit (Qiagen) using a TissueLyser at 25 Hz for a total of 2 minutes, with a 30-second incubation on ice between cycles. The homogenates were centrifuged at 10,000 g for 10 min at 4C  to remove cells and debris, and the Qiagen RNeasy Kit protocol was followed for RNA extraction. On-column DNase treatment was included during RNA isolation. RNA integrity was assessed using a Fragment Analyzer. 1ug total RNA was used on magnetic bead for capture of polyA mRNA.   Illumina Stranded mRNA Prep Kit was used for library preparation according to manufacturers protocol. All samples were sized, quantified, and validated on a Bioanalyzer to ensure high quality for sequencing.