GATA-1 Protein Levels Are Not Altered in CD34⁺ and CD61⁺ Cells, but NF-κb Protein Levels Are Elevated in Patients with Idiopathic Myelofibrosis

GATA-1 and NF-kB are important in megakaryocytopoiesis and pathogenesis of idiopathic myelofibrosis (IMF). Previous studies have shown that GATA-1 levels were normal in mRNA but were lower in protein content in CD61⁺ cells of patients with IMF, and we reported previously that GATA-1 protein levels were not altered in CD61⁺ cells. The current study includes more patients and also measured NF-kB levels. ELISA assay (GATA-1 and NF-kB ELISA kit, Panomics Inc. CA) was used to determine and compare the protein content of nuclear extracts of blood hematopoietic progenitor cells (CD34⁺) and megakaryocytes (CD61⁺ cells) from patients with IMF, patients with other myeloproliferative disorders, and with controls. CD61⁺ cells were obtained from 10-day cultures of blood CD34⁺ cells in serum-free culture medium with thrombopoietin and stem cell growth factor. Our study confirmed that GATA-1 protein levels are not altered in megakaryocytes (cultured CD 61⁺ cells) in patients with IMF (Fig. 1) and NF-kB levels are increased (Table 1). We conclude that GATA-1 protein levels are not altered in patients with IMF compared with controls.

Fig 1.

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GATA-1 Protein levels in CD61⁺ cells Cultured from Patients with IMF & Others

Fig 1.

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GATA-1 Protein levels in CD61⁺ cells Cultured from Patients with IMF & Others

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Table 1. NF-kB Levels (ng/2μg protein)