Figure 1
Live fluorescent imaging of Plasmodium berghei HSP101 during asexual blood-stage development.
(a) Recombination strategy for the endogenous tagging of HSP101. Double crossover integration into the wild-type locus yields recombinant parasites with their endogenous locus tagged by mCherry-3xMyc. For details see Supplementary Fig. S1 and Supplementary Table S1. (b) Intravital competition assay of WT and hsp101-mCherry parasites. Parasite multiplication rates for WT and hsp101-mCherry parasites were 10.7 and 9.6, respectively (non-significant). (c) Western blot analysis of hsp101-mCherry parasites. The predicted size for tagged HSP101 is 133 kDa and was identified correctly using an anti-mCherry antibody. (d) Fluorescent micrographs of live hsp101-mCherry-infected erythrocytes. Shown are representative images of the fluorescent signal of HSP101-mCherry (top), a merge of HSP101-mCherry and cytoplasmic GFP (middle) and differential interference contrast images (DIC, bottom) for three asexual developmental stages. Inset, free merozoite; scale bar, 5 μm.
