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Figure 4: Polyreactive serum anti-PfP0C0 response induced following repeated immunizations with rPfP0 but not BSA-PfP0C0. (a) Sera of mice administered recombinant PfP0 were treated or not with DNP-sepharose beads. Anti-PfP0C0 titre before and after treatment was determined in an ELISA. A representative example of anti-PfP0C0 response of DNP adsorbed (empty circles) and unadsorbed (filled circles) sera. Error bars represent the SEM of three readings. (b) Mice were immunized by four weekly and up to seven monthly injections of rPfP0. Aliquots of the sera were treated with DNP-sepharose beads to adsorb out polyreactive antibodies. The bar graphs depict the percent polyreactivity in sera of mice after multiple immunizations as indicated. Error bars represent the SEM of 3 sera. (c) Box plots showing the 25th and 75th percentiles, together with the median, with whiskers showing the minimum and maximum difference and filled circles representing the outliers, in the ELISA OD responses of DNP unadsorbed (U) and adsorbed (A) sera of all 21 mice immunized with recombinant PfP0; μ (IgM), γ1 (IgG1) γ2a (IgG2a), γ2b (IgG2b), γ3 (IgG3), and α (IgA). Statistically significant values obtained by Paired Wilcoxan signed-rank test are indicated. (d) Autoreactivity of a representative serum sample against bovine insulin (closed circle), dsDNA (open circle), and ssDNA (closed triangle) of mice immunized with rPfP0. Red line represents the response of control unimmunized sera at the lowest dilution (1 : 1000). Error bars represent the SEM of three readings. Similar results were obtained with other sera. (e) Mice were administered BSA-PfP0C0, and the sera were treated or not with DNP-sepharose beads. Anti-PfP0C0 titre before and after treatment was determined in ELISA. A representative example of anti-PfP0C0 response of DNP adsorbed (empty circles) and unadsorbed (filled circles) sera. Error bars represent the SEM of three readings.