Expression of GATA2 and GATA2-AS. (A) Western blots of GATA2 and GAPDH in fibroblasts from the patient (P) and 2 healthy controls (HC1, HC2) showing ∼50% reduction in the ratio of GATA2/GAPDH. Samples were loaded in triplicate for quantification as shown. Three independent experiments were performed with similar results. *P < .05 vs control. (B) Allelic expression was investigated by preparing genomic DNA (gDNA) and complementary DNA (cDNA) from maternal PBMCs, patient PBMCs, and fibroblasts. Two single-nucleotide polymorphisms rs2335237 (exon 1) and rs1573858 (exon 3), heterozygous in the patient and homozygous in the mother, showed preferential expression of the duplicated maternal allele above the peak heights observed in the genomic DNA sequence chromatogram (asterisks). (C) Quantitative polymerase chain reaction analysis of GATA2 expression in PBMCs and fibroblasts comparing ΔΔCt (GATA2-GAPDH) in 2 controls and the patient using 3 primer pairs for GATA2 exons 1-2, exons 3-4, and the GATA2 anti-sense transcript (AS1). (D) GATA2 and GATA2-AS1 expression determined from RNA sequencing data in PBMCs from the patient and 2 controls. (E) Structure of the duplication showing the position of the GATA2 distal enhancer (asterisk) in relation to the second copy of GATA2 and the potential hybrid transcript of GATA2 and RPN1.