1. Methods applied in addition
Supplemental Figure 1 A
Total cohort: 467 RUNX1 mutated AML
Selected cohort: 163 cases
n = 321
69%
n = 53
11%
n = 93
20%
RUNX1 WT loss
(48: CN-LOH, 5: deletion)
n = 53
33%
n = 56
34%
> 1 RUNX1 mutation
1 RUNX1 mutation
n = 54
33% B
Methods applied
Methods applied in addition
cytogenetics
sequencing of RUNX1
genomic array in all cases with RUNX1 mutation load >50%
mutation analyses of 28 genes
genomic arrays
survival analysis
Supplemental Figure 1: Overview of cases and applied methods. (A) the composition of cases is shown for the total cohort (left panel) and for the selected cohort (right panel) of cases. (B) Applied methods for the total cohort of 467 cases (left column) and for the selected cohort (right column) of cases.

2. Supplemental Figure 2
n = 3
n = 15
2 mutations
3 mutations
4 mutations
n = 75
Supplemental Figure 2: Number of mutations in cases with >1 RUNX1 mutation. (A) the number of cases with 2, 3 and 4 mutations is depicted.

3. + + + + + + Supplemental Figure 3
RUNX1 WT loss +
≥2 additional mutations
3 months
>1 RUNX1 mutation +
≥2 additional mutations
14 months
>1 RUNX1 mutation +
0 or 1 additional mutation
17 months
1 RUNX1 mutation +
≥2 additional mutations
20 months
RUNX1 WT loss +
0 or 1 additional mutation
36 months
1 RUNX1 mutation +
0 or 1 additional mutation
not reached
Overall survival
Supplemental Figure 3: Summary of overall survival. Overall survival (depicted by arrows) was determined in the subgroups of RUNX1 WT loss (dark grey), >1 RUNX1 mutation (grey) and 1 RUNX1 mutation (light grey) in combination with ≥2 additional molecular mutations (dark grey) or <2 additional molecular mutations (light grey).