Clonal progression from diagnosis of PMF to leukemic transformation and death. (A) Dynamic changes in the size of the two mutated clones, JAK2V617F and ASXL1-R693X, in three time points during follow up: at the time of presentation of PMF, during the evolution and in blast-phase. (B) Microsatellite analysis on PB granulocytes at diagnosis (left panel) and blast-phase of MF (right panel) for 4 markers on chromosome 9p flanking JAK2. The positions of microsatellite markers used to identify the common 9pLOH region are shown as vertical lines (C) ASXL1 sequencing in paired samples of cDNA from PB granulocytes showing one nonsense mutation (R693X) in the ASXL1 gene at blast-phase, but not at diagnosis (left panel). This mutation was not detected in the hepatic metastatic tissue of lung cancer (right panel).