Extended Data Fig. 3: Loss of H3K79me2 impairs T cell anti-tumour immunity through STAT5.
From: Cancer SLC43A2 alters T cell methionine metabolism and histone methylation
a, Genotyping for Dot1lf/f and Dot1l−/− mice by PCR. b, Effect of Dot1l knockout on histone marks in T cells. c-e, Gene signature comparison between Dot1l−/− and Dot1lf/f CD8+ T cells. Functionally grouped network of enriched categories was generated for the hub genes and their regulators using ClueGO. Visualization has been carried out using Cytoscape 3.7.1. (c). GSEA plot showed enriched apoptotic gene pathway (d) and impaired TCR signalling pathway (e) in Dot1l−/− CD8+ T cells. f, Effect of DOT1L deficiency on T cell function in MC38 tumour. MC38 cells were inoculated into Dot1lf/f and Dot1l−/− mice. Expression of TNFα, IFNγ, and granzyme B in tumour infiltrating CD8+ T cells was determined by FACS. g, Effect of anti-PD-L1 on tumour growth in Dot1l+/+ and Dot1l−/− mice. h, i, B16F10 cells were inoculated into Dot1l−/− and Dot1lf/f mice. Effect of T cell DOT1L deficiency on tumour growth (h) and T cell viability (i) were monitored. j, Real-time PCR showed Stat5a and Stat5b transcripts in fresh or anti-CD3/CD28 activated Dot1lf/f and Dot1l−/− CD8+ T cells. k, Real-time PCR showed Stat5a and Stat5b transcripts in activated CD8+ T cells cultured with fresh medium, B16F10 tumour supernatants (Sup), or supernatants plus methionine (Sup + Met) for 24 h. l–n, RNA-seq showed the effect of DOT1L inhibitor (SGC0946) on human CD8+ T cells (Database: GSE108694). STAT5A and STAT5B (l) transcripts were quantified in human CD8+ T cells treated with DOT1L inhibitor SGC0946. GSEA enrichment plot showed enrichment of apoptotic gene pathway (m) and defects in T cell receptor related pathways (n) in human CD8+ T cells treated with DOT1L inhibitor. o, p, H3K79me2 ChIP-seq in ENCODE database showing Stat5b promoter in mice (o) and humans (p). Data are mean ± s.e.m. Information on sample sizes, experimental number, times, biological replicates, statistical tests, and P values is available in ‘Statistics and reproducibility’ (Methods).
