Extended Data Fig. 5: Aberrant erythroid differentiation in TP53 mutant AML.

a-b, Analysis of erythroid populations in TP53-sAML PDX models. Gating strategy used to identify CD253a+ and erythroid progenitor cells (“ProgE”) (a) and percentage of each erythroid population in hCD45+ bone marrow cells from PDX models (b). n = 5, bars indicate mean ± s.e.m. c, Percentage of cells expressing erythroid markers after culturing CD34 + TP53-sAML cells in conditions promoting myelo-erythroid differentiation in vitro. n = 4, bars indicate mean ± s.e.m. d-e, Force Atlas representation of a CD34⁺ myelofibrosis (MF) atlas (d; Psaila et al, 2020) and latent-semantic index projection of TP53 multi-hit cells from TP53-sAML patients into the MF cellular hierarchy (e). f, Projection of immunophenotypically-defined MEPs into a diffusion map of the single cells from all 14 TP53-sAML patients (as in Fig.2a). g-j, Expression of a comprehensive erythroid (g,h) and myeloid (i,j) gene score derived from a human haematopoietic atlas (Granja et al, 2019) in AML patients from the BeatAML dataset (g,i) and TCGA (h,j) stratified by TP53 mutational status (BeatAML: n = 329 TP53-WT and n = 31 TP53-mutant; TCGA: n = 140 TP53-WT, n = 11 TP53-mutant). k, Expression of a TP53-target gene score using the same p53 target genes as in Extended Data Fig. 4c in patients with high (above median) and low (below median) erythroid scores. l, GATA1/CEBPA gene expression in AML patients from the BeatAML dataset stratified by TP53 mutational status. In (g-l), boxplots represent median, first and third quartiles, and whiskers correspond to 1.5 times the interquartile range. “p” indicates two-sided Wilcoxon rank sum test p-values. m, Erythroid score (left) and CEBPA/GATA1 gene expression ratios (right) in MOLM13 TP53-mutant isogenic cell lines (Boettcher et al, 2019). Boxplots represent median, first and third quartiles, and whiskers correspond to 1.5 times the interquartile range. “p” indicates two-tailed unpaired t-test p-value. n, Fold-change TP53 expression in CD34⁺GFP⁺ MPN primary cells following transduction with a lentiviral shRNA vector targeting TP53 compared to a scramble control (shCTR). n = 3 patients, 3 independent experiments. Barplot indicates mean ± s.e.m. and “p”, two-tailed unpaired t-test p-value.