Generation of siglec-E–deficient mice. (A) Schematic representation of Siglec-E locus, the gene targeting vector, and the predicted mutated Siglec-E gene. (B) Targeting vector to introduce R126D mutation in exon 1 (that contains the sialic acid binding site), with the aim of generating a full-length protein that lacks the ability to bind sialic acid. (C) Lysates from bone marrow neutrophils or bone marrow–derived macrophages cultured for 3 days in the presence of 100 ng/mL LPS to induce siglec-E were immunoprecipitated with sheep anti-mouse siglec-E IgG and subsequently immunoblotted with sheep anti-mouse siglec-E antiserum followed by rabbit anti-sheep IgG-HRP conjugate. *An upper nonspecific band and a lower heavy chain band were common to all samples.