Controlled malaria infection challenge was performed via bite of Anopheles stephensi mosquitoes infected with Pb-PfSPZ a P. berghei parasite strain that expresses P. falciparum CSP (Espinosa et al., 2017). Also, the parasites express GFP or mCherry, allowing infected mosquitoes to be visually identified via microsopy, and 5 positive mosquitoes were sorted onto separate containers. These mosquitoes were fed with sucrose for the first 6 hours and then with water solution a day before the challenge. The following day, mice were anaesthetised and placed on top of the containers to allow the mosquitoes to blood feed for 30 minutes. In some experiments, 42 hours post mosquito bite, mice were euthanised via cervical dislocation and liver was collected, washed twice in PBS, and homogenized in 4 mL denaturing working stock. In other experiments mice were monitored daily for the presence of parasites in the blood which was detected via flow cytometry of blood collected via tail nick. In this case a mouse was considered patent in the day the proportion of RBCs that were GFP+ exceeded 0.1%.

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