Fig. 2.
Glut3 specifically enhances the protumor response of tumor-infiltrating Tregs and modulates their transcriptomic landscape. A B16F10 melanoma growth in Slc2a3+/+Foxp3IRES-Cre (WT) and Slc2a3f/fFoxp3IRES-Cre (KO) mice. B CD8+/CD4+ T-cell ratios in the spleen (Sp) and tumors (TIL) on day 23 postinoculation in the mice described in (A). C, D Percentages of proliferating (Ki67+) non-Treg CD4+ (C) and CD8+ (D) T cells in the spleens and tumors of the mice described in (A). E Frequency of IFNγ-producing splenic and tumor-infiltrating (TIL) CD8+ T cells in the mice described in (A). F, G Gene ontology analysis of B16F10 melanoma TIL CD8+ T cells and CD4+Foxp3- cells in KO mice. H Differential gene expression (DGE) of select immune response genes in KO TIL CD8+ and CD4+Foxp3- T cells. I Representative FACS plot (top) and frequency (bottom) of splenic and B16F10 melanoma TIL-Tregs in WT and KO mice. J Overall change in gene expression in control and tumor-bearing (TBM) splenic and TIL-Tregs upon Slc2a3 deletion. The numbers within the bar graphs represent the total number of genes affected. K Changes in the expression of representative tumor-infiltrating Treg (TITR) signature gene set genes in the spleen and TIL-Tregs of the KO mice compared with the WT mice. L CDF analysis of TITR signature gene set expression in the spleen and TIL-Tregs of the KO mice. M Heatmap depicting the expression of select NF-κB target genes in TIL-Tregs from WT and KO mice. N KEGG pathway analysis of genes downregulated in KO TIL-Tregs. O CDF analysis of NF-κB target gene set expression in splenic and TIL-Tregs from KO mice compared with WT mice. Changes in the expression of select NF-κB target genes specifically downregulated in KO TBM splenic Tregs. The results are pooled from three (A) or two (B–E, I) independent experiments or are from three pooled samples (J–P). The data are presented as the means ± SEMs (A) and means ± SDs (B–E, I). NE, not expressed. ns, nonsignificant, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001; P values are derived from two-way ANOVA (A), unpaired two-tailed Student’s t-test (B–E, I), the Wald test (DESeq2) (H, K, P), and the Kolmogorov‒Smirnov test (L, O)