Immunostaining for serglycin in bone marrow cells from wild-type and serglycin knockout mice and detection of 35S-labeled macromolecules. (A-B) Immunocytochemistry for serglycin; bar, 20 μm; neutrophils and precursor cells from wild-type mouse (A) and serglycin knockout mouse (B). (C) Western blotting of 1 ng recombinant murine serglycin per lane; detection with rabbit anti–murine serglycin IgG, blocked with 0 to 100 ng/mL murine serglycin as indicated above each lane. As seen, the immunoreactivity of the antibody is blocked in a dose-dependent manner by addition of the antigen in solution to the antibody. (D) Immunoelectron microscopy showing serglycin immunoreactivity (10 nm gold particles) in a myeloid cell from wild-type bone marrow; Golgi stacks are indicated by arrows. (E-F) Biosynthesis of 35S-labeled macromolecules in bone marrow cells and SDS-PAGE of cell lysates with (+) or without (−) cABC treatment. (E) Murine bone marrow cells from wild-type (+/+) and serglycin knockout mice (−/−). (F) Myeloid cells from human bone marrow and peripheral blood separated according to maturational stage; the designation of cell populations indicates the major cell types in each group (PMN, peripheral blood neutrophils; BC+SC, band cells and segmented neutrophils; MC+MM, myelocytes and metamyelocytes; MB+PM, myeloblasts and promyelocytes).
