Flow cytometric acquisition of macrophages and confirmation of alternative activation in WT B malayi–implanted macrophages. (A) Gating strategy used to obtain pure macrophage populations as shown with 1 representative WT B malayi–infected individual. After the removal of dead cells, B220, SiglecF, and CD4-positive cells were excluded. F4/80 high cells were then selected and doublets removed based on forward scatter width (FSC-W)/forward scatter area (FSC-A). (B) Intracellular cytokine staining for RELMα expression in thioglycollate-elicited macrophages (top) and B malayi–elicited macrophages (bottom). The scatter profile and macrophage gates on the left refer to 1 representative WT B malayi–infected individual. (C) Bar chart showing percentage of RELMα-positive macrophages from analysis. (B) n = 9 per group. ***P < .001.