Fig. 2: Functional and genetic characterization of peripheral blood samples from the two female carriers of the PGK1 mutation.
A Enzymopathies-related enzyme activity levels of phosphoglycerate kinase 1 (PGK1), glucose-6-phosphate dehydrogenase (G6PD), hexokinase (HK), and pyruvate kinase (PK) were quantified. This table also shows the number of reticulocytes in the propositus (II.1) and her mother (I.2). Note that the PGK1 activity in the propositus (near the lower limit of the normal range), together with the severe reticulocytosis, supports a PGK1 deficiency. B Osmoscan profiles in the mother (I.2) and the propositus (II.1) using a Laser-Assisted Optical Rotation Cell Analyzer (LoRRca). The osmotic gradient ektacytometry (OGE) reflects the deformability of erythrocytes measured in a suspension medium with constant shear stress and a variable osmolality. Informative parameters of this curve are shown in Table S2. C CALRins5 identification, quantification of VAF and mRNA levels of mutated and wild-type PGK1 alleles in the CD3+ and CD15 + PB population of the mother (I.2) and the propositus (II.1). D X-chromosome inactivation ratio (269:279) by HUMARA assay in CD3+ and CD15 + PB maternal (I.2) and propositus (II.1) populations. The following reference limits were established: normal, <80:20; moderate deviation, 80:20–90:10; and marked deviation, >90:10. E Methylation studies of unmanipulated genomic DNA from myeloid progenitors. The CpG island of the PGK1 promoter is shown in green. Methylated bases are shown in red, while unmethylated bases are shown in blue. Single nucleotide variant (SNV) positions are marked in blue. The yellow line indicates the position of the PGK1 p.Ser320Asn variant. The two alleles differentiated by phasing are indicated. PGK1 p.Ser320Asn variant is detected in allele 1.
