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. 2018 Feb 15;8:3085. doi: 10.1038/s41598-018-21551-2

Figure 4.

Figure 4

Parasite sequestration in the brain differs between Mock-control and one-shot RAS-immunized mice after iRBC challenge. C57BL/6 J mice were immunized with one-shot of 3 × 104 RAS (n = 12) or salivary gland debris (Mock; n = 8) and challenged 6 weeks later with 105 iRBC of PbGFP Luccon WT parasites. (A) One-shot RAS-immunized and iRBC-challenged animals do not develop ECM. (B) Delayed blood-stage growth in one-shot RAS-immunized animals compared to Mock-control mice after a challenge with 105 iRBCs. For statistical analysis, a Mann-Whitney Test was applied; day 5, P = 0.002; day 6, P = 0.002 and day 7, P = 0.0002). (C) From day 7 to day 9 after infection, one-shot RAS-immunized mice showed a significantly reduced brain parasite load after a challenge with 105 iRBCs. Groups of mice were sacrificed at the day when Mock-injected mice showed ECM (day 7 after challenge; n = 7 for Mock-injected mice and n = 5 for one-shot RAS-vaccinated mice). For RAS-vaccinated animals, additional groups of mice were sacrificed at day 8 (n = 4) and day 9 (n = 3) after challenge. Bioluminescence was recorded as total flux (photons (p)/second (s)). Statistical analysis was performed using Kruskal-Wallis with Dunn’s Multiple Comparison Test (***p < 0.001; **p < 0.01; ns, not significant). (D) Ex vivo bioluminescence images from RAS and Mock animals at day 7 after challenge. Note the increased bioluminescence signal in ECM-affected Mock animals, which is notable particularly in in the olfactory bulb.