Panel A: A vertical bar graph comparing the ratios of different cell types among live C D 45 positive cells in co-cultures of P S M B 10 G 209 R versus P S M B 10 W T. The x-axis labels are C D 4 positive C D 8 positive, C D 3 positive, and C D 3 positive T C R alpha beta positive. The y-axis shows the ratio P S M B 10 G 209 R over control. The graph includes three data series represented by different colors: blue for Experiment 1, red for Experiment 2, and green for Experiment 3. The dashed line indicates a ratio of 1. Panel B: Flow cytometry pseudo-color plots displaying the proportions of differentiating cells at various stages of T cell differentiation. The plots are divided into two rows: the top row for control cells and the bottom row for P S M B 10 G 209 R cells. Each plot is labeled with specific markers such as C D 34, C D 7, C D 1 a, C D 4, C D 8, C D 3, and T C R α β. The plots show the distribution of cells at different stages of differentiation, including Pro T 1, D N stage, D P stage, C D 3 stage, C D 3 positive T C R alpha beta positive stage, and gated on C D 3 positive cells.
Ex vivo differentiation of CD3+TCR+ cells from PSMB10 WT and PSMB10 G209R HSCs in ATOs. (A) Histograms showing the ratios of DP CD4+CD8+ cells, CD3+ cells, and CD3+TCRαβ+ cells among live CD45+ cells in cocultures of PSMB10 G209R versus PSMB10 WT, based on three independent experiments (see also Fig. 7, A and B). The dashed line indicates a ratio of 1. (B) Flow cytometry pseudocolor plots displaying the proportions of differentiating cells (early T cell progenitor markers CD5, CD7, CD1a, and late T cell differentiation markers CD4, CD8, CD3, TCRαβ) from patient- and healthy donor–derived control cells (gated on live CD45+ cells) in an ATO model performed in a second laboratory. Consistent with results shown in Fig. 7, A and B, there is no differentiation arrest, and mature CD3+TCR+ T cells develop from both PSMB10 WT and PSMB10 G209R HSCs.