Patient characteristics associated with neutropenia. (A-B) Representative patterns of neutrophil (blue line) and platelet (orange line) cycling in the CyN-AML patient during G-CSF treatment. ANCs and platelet counts were plotted for 25 days (left) and for 35 days (right), accordingly. (C) Genetic characterization of double de novo ELANE mutations in the CyN-AML patient. The blue arrows above the Sanger sequencing traces indicate the positions of the mutations. Cloning of a genomic DNA fragment spanning ELANE c.697G<C and c.703delG showed that both mutations were present on the same allele. Two representative examples of sequenced clones are shown. Sequencing of ELANE exon 5 revealed no ELANE mutations in parental DNA isolated from peripheral blood MNCs. (D) Western blot analysis of ELANE protein expression in BM polymorphonuclear cells of the CyN-AML patient and healthy donors. β-actin was used as a loading control. The level of ELANE protein expression was lower in BM polymorphonuclear cells of the CyN-AML patient as compared with healthy donors. The blot shown is representative of 3 independent experiments. BM, bone marrow; MNC, mononuclear cell.