Fig. 3: Genetic profiling of + 9p patients by means of colony and single cell analysis.

A–C ddPCR analysis of the JAK2 mutational burden and copy number on DNA from single CD34+ -derived colonies from +9p patients (n = 139 colonies, with an average of 17 colonies/patient across 8 patients). A Barplot depicting the frequency of colonies split by the number of JAK2 mutated alleles and 9p ploidy. B Jitter plot displaying the distribution of JAK2 mutated alleles and copy number at the single-colony level. C Graphical representation of molecular events’ acquisition order of patient #4, as inferred from ddPCR analysis (n = 15 colonies). Percentages indicate the frequency of colonies carrying the above-depicted 9p ploidy and JAK2 genotype. D–F Single-cell genomic analysis of CD34+ cells and PBMCs from patient #1. The Copy Number Variation profiling of each clone, listed in the rows, at the amplicon level for the entire genome (D) or JAK2 locus (E), is reported in a heatmap. The shades of color depict the ratio between the median amplicon ploidy of a given clone and the one of the diploid reference, which was set to 2. The Wild-Type clone (WT) was set as diploid reference. F Jitter plot displaying the distribution of JAK2 mutated alleles and copy number at the single-cell level (n = 5617 cells). C Was created with Biorender.com.