<i>In vivo</i> erythrophagocytosis assay and <i>ex vivo</i> RBC hemolysis experiment.

<p>A) <i>Ex vivo</i> fluorescent microscopy picture of erythrophagocytozing cell in the spleen of <i>T</i>. <i>b</i>. <i>brucei</i> infected (day 6 p.i.) mice. Left panel: co-culture of spleen cells with unlabeled RBC. Right panel: co-culture of spleen cells with pHrodo-labeled RBC. B) Histogram of PE signal of liver macrophages exposed <i>in vivo</i> to unlabeled RBC (grey tinted area) and pHrodo – labeled RBC (single black line). The delta MFI is the difference between the median fluorescent intensities of the two cell populations. D&E) <i>In vivo</i> erythrophagocytosis assay: delta median fluorescent intensity of myeloid cells in liver (C) and spleen (D) of <i>T</i>. <i>brucei</i> infected (day 6 p.i.) mice. Values represent mean ± SEM of 6 mice per group. E) Relative contribution to phagocytosis: cell numbers were multiplied by the deltaMFI observed for each subpopulation. The surface of the diagram of IFNγR-/- mice displays the total number of phagocytic cells and is calculated relatively to the surface of the diagram of C57BL/6 mice. F) <i>In vitro</i> stimulation of naïve monocytes derived from IFNγ-/- mice with IFNγ induces differentiation to monocyte-derived macrophages. Values represent mean +/- SD of 6 mice per group. G) <i>In vitro</i> erythrophagocytosis assay to determine the effect of IFNγ stimulation on naïve spleen cells from IFNγ-/- mice. Values represent mean ± SEM of 6 mice per group. One representative of 2 independent experiments is shown. *: p-value < 0.05; **: p-value < 0.01; ***: p-value < 0.005; ****: p-value < 0.0001 and if nothing is mentioned the differences were not significant.