Fig. 5. Myc activation reduces immunogenicity in Nras^G12D-driven AML.

A Enrichment of genes correlating with down-regulation of Nras signaling in immunoedited (IE) Nras^G12D AML, as determined from RNA-sequencing of GFP⁺ AML cells isolated from either immunocompetent WT (IE, n = 5) or immunodeficient Rag2^−/−γc^−/− (N-IE, n = 5) recipients. B Relative quantification of Nras copy number in genomic DNA by qPCR in N-IE (n = 5) and IE (n = 5) Nras^G12D AML cells, expressed fold change to the N-IE mean. Each point represents a biologically independent animal, data are presented as mean values +/− SD. C Enrichment of genes correlating with upregulation of MYC transcriptional targets in IE Nras^G12D AML. D Correlation between the relative enrichment of a gene set containing core MYC transcriptional targets³⁶ and genes associated with cytolytic infiltrate in AML³⁷, using bulk expression data from NRAS mutant AML patients²⁹. E Flow cytometry analysis of cell surface expression of immune modulatory molecules on N-IE Nras^G12D AML cells transduced with either a Myc expression construct (MYC) or empty vector (EV) and passaged in Rag2^−/−γc^−/− recipients (mCherry⁺ GFP⁺ CD11b⁺ splenocytes from 5 independent Rag2^−/−γc^−/− recipients) (p = 0.0005 (H2-D^b), p = 0.005 (H2-K^b), p = 0.0001 (MHC Class II), p = 0.0002 (PD-L1), p = 0.0079 (CD86)). Data are presented as mean values +/− SD. F Survival of Rag2^−/−γc^−/− and WT secondary recipients transplanted with 60,000 N-IE Nras^G12D/EV or MYC AML cells. Mantel-Cox test for comparison of Kaplan–Meier curves. (n = 5 per group, p = 0.0027 (EV Rag2^−/−γc^−/− vs EV WT), p = 0.0027 (MYC Rag2^−/−γc^−/− vs MYC WT), p = 0.0027 (EV Rag2^−/−γc^−/− vs MYC Rag2^−/−γc^−/−), p = 0.0133 (EV WT vs MYC WT)) (F). Unpaired two-tailed t-test (E) with Welch’s correction (B). Two-tailed Pearson’s test for association (D). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source Data file.