Fig. 6: Flexible immunogenicity of myeloid tumors.

a Experimental scheme used in b–g. Mouse bone marrow progenitor cells were transduced with MLL-AF9 and were transplanted into sublethally irradiated WT mice. GFP⁺ MLL-AF9 cells were collected from moribund mice (MLL-AF9-2021) and were then transplanted into WT and Rag2^−/− mice (b). Some MLL-AF9-2021 cells were subjected to serial transplantation with non-irradiated recipient mice to establish aggressive AML cells with strong repopulating ability (MLL-AF9-2022). The MLL-AF9-2022 cells were then transplanted into WT and Rag2^−/− mice (c). Figures were created with BioRender (https://app.biorender.com/). b (Left) Kaplan–Meier survival curves are shown. (n = 6 for each group, male mice). **P < 0.01; log-rank test. (Right) Number of GFP⁺ MLL-AF9-2021 cells in peripheral blood of WT and Rag2^−/− mice 14 days after transplantation. Data are shown as means ± s.e.m. (WT: n = 7, Rag2^−/−: n = 8, male mice). ***P < 0.001; two-tailed Mann–Whitney test. c (Left) Kaplan–Meier survival curves are shown. (n = 7 for each group, male mice). (Right) Number of GFP⁺ MLL-AF9-2022 cells in peripheral blood of WT and Rag2^−/− mice 12 days after transplantation. Data are shown as means ± s.e.m. (n = 7 for each group, male mice). *P < 0.05; two-tailed Mann–Whitney test. d MLL-AF9-2021 and MLL-AF9-2022 cells were incubated with DS-5272 at the indicated concentration for 48 h. Cell viability assays were performed using the Cell Counting Kit-8. Data are shown as means ± standard deviation (SD) from three technical replicates. e Multi-Dimensional Scaling of RNA-Seq data. n = 4 per group, male mice). f, g GSEA for down- (f) and up- (g) regulated genes in MLL-AF9-2022 cells compared to MLL-AF9-2021 cells using the Hallmark collections of the GSEA MSigDB (http://software.broadinstitute.org/gsea/msigdb). The x-axis shows the P value (−log10).