Hoechst and JC-1 staining of uninfected and infected blood. In one experiment blood samples were collected from uninfected (A) and P. chabaudi adami DS infected (B) mice. In another experiment blood was collected from uninfected (C) and P. berghei ANKA infected (D) mice. Blood samples were stained with JC-1, anti-CD45 APC eFluor780, anti-CD71 PerCP eFluor710, and Hoechst 33342. Note that plots are a typical representation of results and instrument voltages were slightly different between experiments. Samples were gated up to G3 as in Figure1. A blood sample from a P. chabaudi adami DS infected mouse at approximately 1% parasitaemia was split into three aliquots and serially diluted with uninfected blood to create a dilution curve (E). Calculated parasitaemia was estimated based on the parasitaemia of the undiluted sample, determined by light microscopy, and the dilution factor. Measured parasitaemia for Hoechst was calculated as Q1 + Q2, for JC-1 it was calculated as Q2 + Q3 and for Hoechst + JC-1 it was calculated as Q2 alone (D). Error bars represent SEM for the three replicates.