Figure 5

Functional assays on FPD-AML cell lines. A. The results of a BrdU proliferation assay are indicated for each cell line. Dark bars indicate affected individuals. The standard errors of two independent replicates are shown. A two-way ANOVA resulted in a significant p-value (p < 0.001) between affected and unaffected individuals. B. Examples of the tubulin polymerization assay for an affected and an unaffected individuals in each family. s:soluble tubulin; p:polymerized tubulin. C. The percentage of polymerized tubulin is shown for each cell line. Dark bars indicate affected individuals. The standard errors of three independent replicates are indicated. A two-way ANOVA resulted in a significant p-value (p < 0.002) between affected and unaffected individuals. D. Percentage of polymerized tubulin in the same cell lines before (darker left bars) and after (second bars) induction of polymerization by Taxol. A significant smaller induction is observed in affected individuals (dark bars) as demonstrated by an ANOVA (p < 0.0003). E. Glycophorin A assay. The numbers of N0 (loss of the M allele), NN (mutation changing M to N allele) or total mutant (both N0 and NN) cells are indicated for each individual. The standard errors of three to five technical replicates are indicated. Dark bars represent affected individuals (A1-A2). The control C5 is the unaffected sister of patient A1. ANOVAs were performed for each kind of mutation and the p-values are indicated.