Fig. 6: Gata1 or Xbp1 ablation prevents AML246 maturation into SSC^HIGH eosinophil-like cells.

a Strategy for CRISPR/Cas9-mediated gene knockout in AML246-Cherry cells. b Flow cytometry of bone marrow from representative Rag1^−/− mice transplanted with the indicated AML246 single cell clones, allowed to develop disease, and then treated with Dox for 8 days (Table S3). A control sgRNA clone (top row) is compared to two independent AML246-Cherry Gata1 KO clones (second and third rows) and two independent AML246-Cherry Xbp1 KO clones (fourth and fifth rows). Data are representative of two independent experiments with similar results. Cytospins of sorted mCherry⁺ AML-derived cells are also shown. c Proportion of mCherry⁺ AML cells in the peripheral blood of mice transplanted with AML246-Cherry control sgRNA clones (black), Gata1 knockout clones (red), or Xbp1 knockout clones (blue) upon disease establishment when Dox treatment commenced. Mean +/− SD. p = 0.21 and p = 0.31 for Gata1 and Xbp1 ko clones respectively versus control, unpaired two-sided Student’s t-test. n = 5 mice for control sgRNA clones and Gata1 knockout clones, and n = 4 mice for Xbp1 knockout clones over two independent experiments. d Proportion of mCherry⁺ AML cells that were also SSC^HIGH in the bone marrow and spleen of mice from c following 8 days Dox treatment. Mean +/− SD. Gata1 versus control *p = 0.017 and p = 0.047 for bone marrow and spleen respectively, Xbp1 versus control *p = 0.019 and p = 0.037 respectively, unpaired two-sided Student’s t-test. Mouse numbers as for 6c.