Figure EV3. PyLARC2 immunization of elicits cross-species humoral responses against P. berghei liver and blood stages.

(A) Plasma was collected from six PyLARC2 immunized SW mice and pooled. The pooled plasma was then diluted 1:100 and used to evaluate reactivity of immune sera to antigens at different timepoints of P. berghei ANKA WT LS development at 36 hpi using IFA. Sera is shown in red, anti-mHSP70 antibody is used as control (green) and DNA is stained with DAPI (blue). Scale bar is 20 µm. In more mature P. berghei LS, PyLARC2 IgG displayed cross-reactivity to P. berghei antigens located in both the cytoplasm and the membranes of the parasite. No reactivity was observed in plasma harvested from mock-immunized mice, indicating that the staining was specific to the PyLARC2 immunized mice. (B) Plasma was collected from six PyLARC2 immunized SW or BALB/cJ mice. Pooled plasma for each mouse strain was diluted 1:100 and used to evaluate reactivity of immune sera to antigens expressed in P. berghei ANKA WT blood stages. Sera is shown in red, anti-MSP1antibody is used as control (green) and DNA is stained with DAPI (blue). Scale bar is 10 µm. Immune sera from both SW and BALB/cJ mice reacted with antigens expressed in the cytoplasm of blood stages. No reactivity was observed in plasma harvested from mock-immunized mice, indicating that the staining was specific to immune sera from PyLARC2 immunized mice.