Figure 2: Mutational architecture and subclonal evolution of SF3B1-mutated MDS-RS bone marrow cells.

Panels show the gene mutations with their MABs in primary CD34⁺ bone marrow, HEC cells and LTC samples from each patient. Outer red circle represents the total SF3B1 clone and the inner sections of the circle represents the SF3B1 subclones. MABs for HEC samples are the average of all mouse samples (where applicable, that is, >1 mouse). Percentages displayed in the pie chart indicate the MAB frequency modulation in HEC and LTC samples versus CD34⁺ day 0. *P<0.01% (t-test) for difference in SF3B1 MAB frequency between CD34⁺ day 0 and HEC and/or LTC samples for the MDS3 sample. For more details about statistical analysis of the variation in MAB frequencies, refer to Supplementary Fig. 5. The table on the right side of each panel represents the mutation status of the respective genes in the HSC, MPP, GMP and MEP cell populations for each patient. Samples used to isolate cell fractions (HSCs, MPPs, GMPs and MEPs) were taken ∼18 months later than the first sample used to isolate CD34⁺ cells. Three independent PCRs were performed to confirm/determine the MAB throughout the experiments. In the tables on the right side, the black boxes indicate that MAB for gene mutations was the same as the primary CD34⁺ cells, dark grey boxes show MAB for gene mutations that was≤2-fold as compared to the primary CD34⁺ cells, light grey boxes represent MAB for gene mutations that was ≥2-fold as compared with the primary CD34⁺ cells and pink boxes indicate not detected or below background noise level. MAB mutant allele burden; HEC, human engrafted cell; LTC, long-term culture; BM, bone marrow; Mut, mutation.