JAK2V617F induces centrosome and chromosome abnormalities. (A) Centrosome abnormalities in mitosis. Examples of Ba/F3-JAK2V617F cells with 2, 3, or 4 centrosomes visualized by immunofluorescence with an anti–γ-tubulin antibody (top panels). The bottom panels illustrate the corresponding Hoechst staining showing the cell nucleus. (B) The percentage of cells in metaphase with extra centrosomes (> 2) was measured in Ba/F3-expressing JAK2wt or JAK2V617F cells or in parental cell lines. At least 200 cells in metaphase were counted, and significant difference was analyzed using χ2 tests. *Significant differences (P < .001) compared with Ba/F3 control cell lines. (C) The ploidy of the different cell lines was analyzed. At least 100 cells in metaphase per cell line were analyzed, and a nonparametric analysis of variance was used for statistical significance of the distribution. Significant difference (P < .01) was found in Ba/F3 expressing JAK2wt and JAK2V617F compared with Ba/F3 control cell lines. (D) Histograms represent the percentage of cells with 39 chromosomes and less or more than 39 chromosomes in Ba/F3 cell lines. (E) An example of metaphase cells spread from Ba/F3-EPOR cells displaying SCEs. Then, SCEs were counted on cells in metaphase in each cell line. At least 50 cells in metaphase were counted, and a nonparametric analysis of variance was used for statistical significance of the distribution. Significant difference (P < .001) was found in Ba/F3 expressing JAK2V617F compared with Ba/F3 control cell lines.
