Journal: Cell metabolism

Article Title: The glucose transporter GLUT3 controls T helper 17 cell responses through glycolytic-epigenetic reprogramming.

doi: 10.1016/j.cmet.2022.02.015

Figure Lengend Snippet: Figure 1. GLUT3 is required for the effector function of Th17 cells (A) Immunoblot analysis of murine GLUT3, ACLY, IRF4, NFATc1, and GAPDH expression. (B) Analysis of Slc2a3 (GLUT3) gene expression in naive CD4+ T cells and T helper (Th) cell subsets by qRT-PCR; mean ± SEM of 5–6 mice. (C and D) Glycolytic proton efflux rate (glycoPER) analyses of WT and GLUT3-deficient Th1 (C) and Th17 (D) cells using a Seahorse extracellular flux analyzer; mean ± SEM of 5 mice. (E) Proliferation analysis of WT and GLUT3-deficient Th1 and Th17 cells.

Article Snippet: After an 1 h incubation at RT with mouse-anti-Tom20 (Abcam, clone EPR15581-39) and rabbit-anti-human GLUT3 (BioRad), the samples were washed and stained for 1 h with secondary donkey anti-rabbit IgG (H+L) Alexa Fluor 488 and donkey anti-mouse IgG (H+L) Alexa Fluor 647 antibodies (both Invitrogen).

Techniques: Western Blot, Expressing, Gene Expression, Quantitative RT-PCR

Journal: Cell metabolism

Article Title: The glucose transporter GLUT3 controls T helper 17 cell responses through glycolytic-epigenetic reprogramming.

doi: 10.1016/j.cmet.2022.02.015

Figure Lengend Snippet: Figure 2. Ablation of GLUT3 in T cells prevents autoimmunity (A–D) Slc2a3fl/flCd4Cre mice are protected from experimental autoimmune encephalomyelitis (EAE). (A) Clinical EAE scores of WT and Slc2a3fl/flCd4Cre mice after immunization with MOG35-55 peptide emulsified in CFA; mean ± SEM of 9 mice per cohort.

Article Snippet: After an 1 h incubation at RT with mouse-anti-Tom20 (Abcam, clone EPR15581-39) and rabbit-anti-human GLUT3 (BioRad), the samples were washed and stained for 1 h with secondary donkey anti-rabbit IgG (H+L) Alexa Fluor 488 and donkey anti-mouse IgG (H+L) Alexa Fluor 647 antibodies (both Invitrogen).

Techniques:

Journal: Cell metabolism

Article Title: The glucose transporter GLUT3 controls T helper 17 cell responses through glycolytic-epigenetic reprogramming.

doi: 10.1016/j.cmet.2022.02.015

Figure Lengend Snippet: Figure 3. GLUT3 controls a complex metabolic-transcriptional network in Th17 cells (A) Principal component (PC) analysis of WT and GLUT3-deficient (Slc2a3fl/flCd4Cre) Th1 and Th17 cell RNA-seq data; n = 3 biological replicates per T cell subset and genotype. (B and C) MA plots of differentially expressed genes (DEGs) in WT versus GLUT3-deficient Th1 (B) and Th17 cells (C); genes significantly (p adjusted < 0.01) upregulated and downregulated are depicted in red and blue, respectively. (D) Venn diagram analyses of >4-fold DEGs (p adjusted < 0.01) of GLUT3-deficient Th1 and Th17 cells. (E) Gene set enrichment analysis (GSEA) of WT versus GLUT3-deficient Th17 cells. (F) Network clustering of significantly (p < 0.005) enriched gene expression signatures to identify dysregulated physiological processes in GLUT3-deficient Th17 cells. Downegulated and upregulated gene sets in GLUT3-deficient Th17 cells compared with WT are shown in blue and red, respectively. (G) Heatmap analysis of selected genes in GLUT3-deficient and WT Th1 and Th17 cells. (H) GSEAs of WT versus GLUT3-deficient Th17 cells highlight impaired mitochondrial gene expression and function.

Article Snippet: After an 1 h incubation at RT with mouse-anti-Tom20 (Abcam, clone EPR15581-39) and rabbit-anti-human GLUT3 (BioRad), the samples were washed and stained for 1 h with secondary donkey anti-rabbit IgG (H+L) Alexa Fluor 488 and donkey anti-mouse IgG (H+L) Alexa Fluor 647 antibodies (both Invitrogen).

Techniques: RNA Sequencing, Gene Expression