Generation of myeloid-specific PDI CKO mice. (A) Targeting construct and homologous recombination. (B) Southern blotting analysis of genomic DNA isolated from mouse tails. (C) Polymerase chain reaction analysis of WT, heterozygous, and CKO mice with primers for floxed PDI and Lys-Cre. (D) Immunoblotting with lysates of cells isolated from WT (W), heterozygous (H), and homozygous PDI CKO (K) mice. The band density of PDI represents mean ± SD (n = 5-6 mice per group). (E-J) Flow cytometric analysis shows the expression of β2 integrins, L-selectin, PSGL-1, and CD44 on unstimulated (dot line) and fMLF-stimulated (black line) WT (WT) and PDI CKO (KO) neutrophils. The gray histogram represents the fluorescence intensity of control IgG on stimulated neutrophils. The mean fluorescence intensity of antibodies was normalized to that of control IgG, and data are shown as 100% (mean ± SD, n = 3-6 mice per group). PSGL-1, P-selectin glycoprotein ligand-1.