Graphpad prism histogram6/23/2023 ![]() ![]() ![]() Adjusted p values for all comparisons were calculated to be < 0.0001 (shown as ∗∗∗∗ in the plot) and considered statistically significant. MFI values were compared using the Brown-Forsythe and Welch ANOVA tests followed by Dunnett’s multiple-comparison test (GraphPad Prism, v.9.3.1). The median for each group is depicted by the teal horizontal lines. ![]() Bottom panel shows the median fluorescence intensity (MFI) of the binding of several antisera collected from mice infected with either VSG2 WT (black dots, n = 18) or VSG2 AAA (gray dots, n = 12) trypanosomes to VSG2 WT and VSG2 AAA cell lines (x axis). Unstained cells were used as a negative control (black peaks). (B) Histograms reflecting binding intensities of antisera collected from mice infected with VSG2 WT- or VSG2 AAA-covered parasites (top two panels, respectively) to VSG2 WT and VSG2 AAA cells (blue and red correspond to two independent clones of VSG2 WT parasites green and orange correspond to two independent clones of VSG2 AAA parasites). Shannon entropy shows clonal diversity on a sequence level with a value of 1.0 representing 100% clonal diversity (no clones), while a value of 0.0 corresponding to 0% clonal diversity (only clones). Genes from both chains that appeared only once and resulted in single heavy-light pairings, were considered background and were removed from the plot. The heavy and light chain variable gene pairings that form the antibodies are illustrated as connector lines starting from the heavy chain genes, providing also information about the frequency of the pairings. Different colors depict each heavy chain variable gene (bottom half of the plot), and each light chain variable gene (top half of the plot). (C) Circos plots generated for the VSG2 WT V gene signatures of two independent mice (n = 2) whose infections were treated with diminazene (left, n = 96 pairs) and of two additional mice (n = 2) that naturally cleared the first peak of parasitemia at days 6 (center, n = 49 pairs) and 7 (right, n = 44 pairs) post infection.
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