Generation of mMCP-11–deficient mice. (A) Diagram shows gene targeting for the generation of mMCP-11–deficient mice. (B) Charts show results of flow cytometric analysis of indicated surface markers on bone marrow cells isolated from wild-type (WT) and mMCP-11–deficient (KO) littermate mice. c-kit–negative cells were gated and displayed. (C) Graphs show the numbers of basophils in the bone marrow, spleen, and peripheral blood of WT and KO mice (mean ± standard deviation [SD], n = 3 mice each). (D) Graphs show results of quantitative PCR analysis for the expression of Mcpt11 and Mcpt8 mRNAs in c-kit⁻CD49b⁺CD200R3⁺ basophils isolated from the bone marrow of WT and KO mice. (E) Western blot analysis for the expression of indicated proteins in bone marrow–derived basophils (BMBAs) generated from WT and KO mice. (F-G) BMBAs generated from WT (gray bars) and KO (black bars) mice were sensitized with anti-TNP IgE and then stimulated with TNP-OVA or control OVA. In panel F, the extent of degranulation was examined by using a β-hexosaminidase release assay (mean ± SD, n = 3 wells). In panel G, the concentration of cytokines IL-4 and IL-6 in culture supernatants was measured by using ELISA and bead-based immunoassay, respectively (mean ± SD, n = 3 wells). (H) Migration ability of WT (gray bars) and KO (black bars) BMBAs in response to chemokine MIP-2 (100 ng/mL) or control PBS was examined by using a transwell migration assay. The number of cells that migrated from the upper to lower chamber during 2-hour culture was counted (mean ± SD, n = 3 chambers each). Data shown in panels B-H are representative of at least 3 independent experiments. n.s., not statistically significant.