Figure 1.
Figure 1. Sequence analysis, JAK2V617F/JAK2total, and clonality results for patients with MPDs. (A) The frequency of the JAK2V617F allele as determined by sequencing and the real-time PCR (RT-PCR) assay in MPD patients according to granulocyte clonality, demonstrating that DNA resequencing more frequently identifies JAK2V617F mutations in MPD patients with clonal granulocytes but that RT-PCR is able to detect JAK2V617F mutations in patients with clonal and polyclonal granulocytes. (B) The frequency of the JAK2V617F allele as determined by sequencing and by RT-PCR in PV patients according to granulocyte clonality, demonstrating that DNA resequencing more frequently identifies JAK2V617F mutations in PV patients with clonal granulocytes but that RT-PCR is able to detect JAK2V617F mutations in PV patients with clonal and polyclonal granulocytes. (C) The frequency of the JAK2V617F allele as determined by sequencing and by the real-time PCR assay in ET patients according to granulocyte clonality, demonstrating that DNA resequencing more frequently identifies JAK2V617F mutations in ET patients with clonal granulocytes but that RT-PCR is able to detect JAK2V617F mutations in ET patients with clonal and polyclonal granulocytes. (D) Sequence traces of JAK2 exon 14 along with results from quantitative real-time assays for JAK2V617F and clonality results. The top trace shows a sequence trace from a patient with PV and a homozygous JAK2V617F mutation; the second trace shows a sequence trace from a patient with PV and a heterozygous JAK2V617F mutation; and the third trace shows a sequence trace from a patient with ET who was negative for the JAK2V617F mutation by sequence analysis and RT-PCR; each of these 3 patients had allele skewing consistent with clonal granulopoiesis. The bottom trace shows a sequence trace from a patient with PV who was scored as negative for the JAK2V617F mutation by DNA resequencing; there is a small peak (arrow) suggesting that the JAK2V617F mutation may be present in a subpopulation of cells. RT-PCR confirms that JAK2V617F is present in a minor population of cells, and clonality analysis demonstrates a lack of allele skewing consistent with polyclonal granulopoiesis.

Sequence analysis, JAK2V617F/JAK2total, and clonality results for patients with MPDs. (A) The frequency of the JAK2V617F allele as determined by sequencing and the real-time PCR (RT-PCR) assay in MPD patients according to granulocyte clonality, demonstrating that DNA resequencing more frequently identifies JAK2V617F mutations in MPD patients with clonal granulocytes but that RT-PCR is able to detect JAK2V617F mutations in patients with clonal and polyclonal granulocytes. (B) The frequency of the JAK2V617F allele as determined by sequencing and by RT-PCR in PV patients according to granulocyte clonality, demonstrating that DNA resequencing more frequently identifies JAK2V617F mutations in PV patients with clonal granulocytes but that RT-PCR is able to detect JAK2V617F mutations in PV patients with clonal and polyclonal granulocytes. (C) The frequency of the JAK2V617F allele as determined by sequencing and by the real-time PCR assay in ET patients according to granulocyte clonality, demonstrating that DNA resequencing more frequently identifies JAK2V617F mutations in ET patients with clonal granulocytes but that RT-PCR is able to detect JAK2V617F mutations in ET patients with clonal and polyclonal granulocytes. (D) Sequence traces of JAK2 exon 14 along with results from quantitative real-time assays for JAK2V617F and clonality results. The top trace shows a sequence trace from a patient with PV and a homozygous JAK2V617F mutation; the second trace shows a sequence trace from a patient with PV and a heterozygous JAK2V617F mutation; and the third trace shows a sequence trace from a patient with ET who was negative for the JAK2V617F mutation by sequence analysis and RT-PCR; each of these 3 patients had allele skewing consistent with clonal granulopoiesis. The bottom trace shows a sequence trace from a patient with PV who was scored as negative for the JAK2V617F mutation by DNA resequencing; there is a small peak (arrow) suggesting that the JAK2V617F mutation may be present in a subpopulation of cells. RT-PCR confirms that JAK2V617F is present in a minor population of cells, and clonality analysis demonstrates a lack of allele skewing consistent with polyclonal granulopoiesis.

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