Figure 2.

Monoallelic CD177 mRNA expression in neutrophil-differentiated CD34⁺ HSC and neonatal neutrophils. CD34⁺ HSC were differentiated over 7 d into neutrophils and (A) acquired neutrophil surface markers CD11b and CD18, together with a bimodal CD177 phenotype. A representative flow cytometry analysis from four independent differentiation experiments is shown. Solid lines represent staining with the specific antibodies and dotted lines represent the isotype control. (B) CD177 gene expression increased progressively in CD34⁺ HSCs during 7 d of neutrophil differentiation (qRT-PCR; n = 4 different CD34⁺ HSC donors). (C) Sanger sequencing of genomic DNA (gDNA) and mRNA from CD34⁺ HSC showed a heterozygous AC SNP⁷⁹³ (rs10425835) in gDNA and biallelic CD177 mRNA expression in undifferentiated cells at day 0. The C allele was silenced during neutrophil differentiation up to day 7 (n = 4 different CD34⁺ HSC donors). (D) Analysis of neutrophil-differentiated CD34⁺ HSCs and neonatal neutrophils from the same cord blood indicate an informative heterozygous AC SNP⁷⁹³ in the gDNA from CD34⁺ HSCs.