Molecular profile of subclonal TP53 mutations. (A) Allele frequency of the 85 TP53 mutations identified by ultra-deep-NGS. Mutations are ordered according to their allelic abundance. Mutations that tested positive (clonal mutations: gray bars) and negative (subclonal mutations: red bars) by Sanger sequencing are indicated. (B) Prevalence of TP53 lesions according to their clonal representation in the study cohort of 309 newly diagnosed CLL (for each category, the crude number of patients is represented). (C) Comparison of the molecular profile of subclonal mutations from the CLL study cohort (n = 50) vs clonal mutations from public CLL databases (n = 257; see Zenz et al²⁹ ). p, P values by Fisher's exact test corrected for multiple hypothesis testing. (D) Comparison of the transition/transversion profile between subclonal TP53 substitutions from the CLL study cohort (n = 48) and clonal TP53 substitutions from public CLL databases (n = 210; see Zenz et al²⁹ ). p, P values by Fisher's exact test corrected for multiple hypothesis testing. (E) Schematic diagram of the TP53 protein with its conserved functional domains. Color-coded shapes indicate the position of subclonal TP53 mutations from the CLL study cohort (n = 50; red shapes) and clonal TP53 mutations from public CLL databases (n = 257; gray shapes; see Zenz et al²⁹ ). Hot spot codons recurrently affected by both subclonal and clonal TP53 mutations are highlighted. (F) Residual CDKN1A transactivation capacity of subclonal TP53 missense substitutions from the CLL study cohort (n = 39; red box) vs clonal TP53 missense substitutions from public CLL databases (n = 193; gray box; see Zenz et al²⁹ ). The band inside the box is the median value. The bottom and top of the box are the 25th and 75th quartiles. The ends of the whiskers are the second percentile and the 98th percentile. p, P value by Mann-Whitney test.