Fig. 5

Single-cell sequencing analysis confirms enhanced MDSCs recruitment and chemokines secretion in tumor-bearing Mettl3-cKO mice. Seurat analysis eventually clustered 11 cell populations from both WT and Mettl3-cKO mice. (A) Integrated UMAP image of 11 major cell populations in PerC from tumor-bearing WT mice. (B) Merged UMAP image of two analyzed samples: PerC cells from tumor-bearing WT and Mettl3-cKO mice. (C) Representative proportions of distinct cell populations in WT and Mettl3-cKO mice. (D) The transcriptional level of CD45 mRNA (by Ptprc) among 24 distinct cell populations. (E) Transcriptional expression of different cytokines and chemokines in subpopulations at the single-cell level. (F) Gene Ontology Enrichment Analysis using the clusterProfiler package revealed differentially expressed genes of “Granulocyte” subpopulations between WT and Mettl3-cKO mice. A total of 815 up-regulated and 1631 down-regulated genes were identified in granulocytes from Mettl3-cKO mice compared with that in WT mice. (G) KEGG pathway analysis of differential expressed genes of “Granulocyte” subpopulations between WT and Mettl3-cKO mice. Size is proportional to the number of differential expressed genes. Red to blue colors represent different adjusted P values. (H) The top ten differential expressed genes in granulocytes of Mettl3-cKO mice are listed. (I) Cybb mRNA expression level in granulocytes isolated from tumor-bearing WT or Mettl3-cKO mice was verified by qRT-PCR. (J) Cybb mRNA expression level in granulocytes isolated from the bone marrow of WT or Mettl3-cKO mice was verified by qRT-PCR.