Analysis of clonal markers in patient NA-02. (A) Bone marrow samples at MPD diagnosis. The presence of the JAK2-V617F mutation was determined by AS-PCR using DNA from purified cells: CD15⁺, myeloid cells isolated by FACS; CD3⁺, T cells isolated by FACS. %T indicates the allelic ratio between the mutant and wild-type JAK2 allele. 9pLOH was determined with the microsatellite markers D9S1810 and D9S288 using DNA from purified cells as for JAK2. XCIP was determined by AS-PCR for a G/T polymorphism in the MPP1 mRNA. The relative expression of the 2 MPP1 alleles was determined by comparing the G and T peak intensities obtained by the allele-specific reverse transcription-PCR assay. (B) Peripheral blood samples at AML transformation. GRA, Granulocytes isolated by Ficoll density centrifugation; CD34⁺, leukemic blasts isolated by FACS; CD3⁺, T cells isolated by FACS. The assays were performed as in (A).