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Blood, Vol. 109, Issue 1, 71-77, January 1, 2007
Evidence that the JAK2 G1849T (V617F) mutation occurs in a lymphomyeloid progenitor in polycythemia vera and idiopathic myelofibrosis
Blood Delhommeau et al.
109: 71
Supplemental materials for: Delhommeau et al, Vol 109, Issue 1, 71-77
Files in this Data Supplement:
- Figure S1. Semiquantitative estimation of the JAK2 G1849T–total JAK2 ratio using sequencing and real-time PCR assay (PDF, 1.58 MB) -
100% mutated DNA was mixed in various proportions with 100% normal control DNA. The sequence traces and real-time PCR amplification plots indicating mutated (m) and wild-type (wt) JAK2 amplification plots of each dilution are shown, demonstrating the correlation of the techniques. The sensitivity of the sequencing was 5%, whereas the sensitivity of the real-time PCR assay was about 1% to 2% of mutated allele. However, because the purity of the lymphoid cell populations studied in this work was 98%, results below 2% of mutated allele were arbitrarily considered as negative because 2% of mutated myeloid cells could contaminate the samples. Semiquantitative results were expressed as shown in the right part of the figure.
- Figure S2. Immunophenotype and genotype of CD34+CD38–-derived myeloid, NK/myeloid, and B/NK/myeloid clones from healthy bone marrow, PV bone marrow, and IMF peripheral blood (PDF, 411 KB) -
CD19 PE/CD15 FITC and CD56 APC/CD15 FITC dot plots of myeloid, NK/myeloid, and B/NK/myeloid clones from control and patients are shown. IMF and PV clones were genotyped by sequencing or real-time allele specific PCR (m, JAK2 G1849T; wt, wild-type JAK2): a, sequence trace of an IMF homozygous mutated myeloid clone; b, sequence trace of a IMF heterozygous NK/myeloid clone; c, sequence trace of a IMF wild-type B/NK/myeloid clone; d, amplification plot of a PV homozygous mutated myeloid clone; e, amplification plot of a PV heterozygous NK/myeloid clone; f, amplification plot of a PV wild-type B/NK/myeloid clone.
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