Expression of G193X Elane. (A) The genomic organization of murine Elane is shown in the top panel. The asterisk (*) represents the introduced G193X mutation; an EcoR1 site was just inserted immediately 3′ to this mutation to facilitate genotyping. (B) RT-PCR analysis of bone marrow RNA isolated from wild-type (WT) and mice heterozygous (Het) or homozygous (G193X) for the G193X Elane allele. The genotyping primers are depicted in panel A. After RT-PCR, the amplicons were incubated with EcorR1 (+) or buffer alone (−); EcoR1 cleaves G193X but not wild-type Elane cDNA. (C) Western blot analysis of cell lysates isolated from the bone marrow of wild-type (WT) or Elane^−/− (NE^−/−) mice. (D) Western blot analysis of cell lysates prepared from unfractionated bone marrow cells (whole BM) or cultured granulocytic precursors. Where indicated, cell lysates were treated with PNGase F before analysis. The positions of mature and precursor wild-type NE are indicated with an arrow or arrowhead, respectively. Data are representative of 3 independent experiments.