Supplementary Figure 4: Elmo1 deletion in neutrophils inhibits neutrophil migration.

a) Total bone marrow cells were purified as described in the Methods, and cell migration toward the indicated concentrations of LTB4 or CXCL1 was evaluated after 3 hours at 37 °C by flow cytometry. Each symbol represents an individual animal. b) Transcript levels of the LTB4 receptor Blt1 are analyzed by qPCR. Each symbol represents an individual animal. c) Flow cytometry analysis of cell surface levels of CXCR2 in Elmo1^+/+ and Elmo1^–/– mice on Ly6G⁺ neutrophils in the blood (top panels, n = 9, 11) and the bone marrow (bottom panels, n = 3, 3). MFI, mean fluorescence index. FMO, fluorescence minus one. Each symbol represents an individual animal. d) Flow cytometry analysis of cell surface levels of FcγRIII and FcγRIV on CD11b⁺Ly6G⁺ neutrophils in the blood (top panels) and the bone marrow (bottom panels) of Elmo1^+/+ and Elmo1^–/– mice. e) Immunoblotting of FcγRI protein expression in Ly6G⁺ neutrophils purified from the bone marrow of Elmo1^+/+ and Elmo1^–/– mice. The blot was cropped to show relevant bands in non-adjacent lanes. f) Immunoblotting of Syk protein expression in Ly6G⁺ neutrophils purified from the bone marrow of Elmo1^+/+ and Elmo1^–/– mice. The blot was cropped to show relevant bands in non-adjacent lanes.