PAPA patient macrophages make membrane projections associated with matrix degradation and have enhanced invasive properties. (A) Representative images of gelatin degradation by control and PAPA macrophages on Oregon Green 488 gelatin-coated coverslips (green). Podosomes and membrane projections were stained with rhodamine phalloidin (magenta). Areas degraded by the macrophages appear black in the gelatin image. Arrowheads indicate projections overlaying degraded areas. (B) Quantification of gelatin degraded by control and PAPA macrophages as a percentage of the total cell area. (C) Cell area was determined for all cells in the assay and normalized to the value of the control macrophages. (D-E) Quantification of macrophage chemotaxis to macrophage colony-stimulating factor (M-CSF) in (D) Transwell devices or (E) Matrigel-coated invasion chambers. Chemotactic index is the ratio of cells migrating to M-CSF over medium alone. Representative images of the filters are shown below. Scale bars, 20 μm (A) or 500 μm (D,E). All values are mean ± SEM from 3 independent experiments. *P < .05; **P < .01 as determined by a paired Student t test (C) unpaired Student t test (D,E) or ANOVA with repeated measures using a compound symmetry correlation structure (B).