AML patients from Leucegene cohort carrying germline and somatic RUNX1 mutations. (A) Pipeline used for the identification of germline and somatic RUNX1 mutations. (B) Sequencing chromatograms of leukemic complementary DNA (cDNA)/normal DNA pairs covering mutation sites for RUNX1 and control oncogene. Refer to supplemental Figures 1 and 2 for a complete analysis of leukemic/normal DNA pairs. (C) Primary structure of RUNX1 protein with identified germline mutations (upper panel) and somatic mutations (lower panel). Pie charts show VAF for each mutation, as revealed by RNA-sequencing. Predicted effect of mutation on protein function was determined by prediction tools when not previously described in functional studies and it is depicted by the color scheme. Mutations that have been validated in functional studies are highlighted in bold type and are marked with a pound sign (#); mutations that have been described in RUNX1-FPD pedigrees are highlighted in bold type and are marked with an asterisk (*). Refer to supplemental Table 6 for the references to such studies. Protein domains and mutation positions are based on isoform NP_001745.2. RUNT: 85-206, TAD: 318-398, RUNXI: 389-480. (D) Clinical and genetic characteristics of AML patients with germline and somatic RUNX1 mutations. C, complex karyotype; FAB, French-American-British; IQR, interquartile range; M, male; NK, normal karyotype; PCR, polymerase chain reaction; WBC, white blood cell count.