Extended Data Fig. 4: Regulation of Glut2 expression by HIF1α.
From: The glucose transporter 2 regulates CD8+ T cell function via environment sensing
(a-b) T cells were incubated overnight at 20% or 5% oxygen. Expression of Hif1α was measured by deconvolution microscopy (a) or flow cytometry (b). Representative images and histograms, and mean corrected total cell fluorescence (CTCF, a) or MFI (b) in a representative experiment are shown (±SD; n = 4-5 N = 2). (c) CD8 + T cells were purified from HIF+ or HIF- mice and Hif1α mRNA expression was measured by RT-PCR. Hif1α gene expression was normalized to housekeeping gene tubulin (±SD; n = 4 N = 3). (d-e) Glut2 (d) and Gal-9 (e) expression by 3-day antibody-activated purified naïve CD4+ T cells from Hif1+ or HIF- mice were assessed for surface expression of Glut2 by flow cytometry. Representative histograms and mean data in a representative experiment (±SD; n = 10, N = 3) (f) Glut2 expression by 3-day antibody-activated naïve Gal-9+ or Gal-9- CD4+ T cells was assessed by flow cytometry. Histograms and mean data from a representative experiment are shown (±SD; n = 8, N = 3). (g) Naïve CD4+ T cells were antibody-activated for 3 days in the presence or absence of recombinant Gal-9 (30 nM) with or without the Gal-9 competitive inhibitor lactose (30 mM) before analysing surface expression of Glut2 by flow cytometry. Representative histograms and mean data from a representative experiment are shown (±SD; n = 3, N = 2). (h) Expression of Gal-9 by 3-day antibody-activated CD4+ and CD8 + T cells was analysed by flow cytometry. The mean data of a representative experiment performed in triplicate is shown (±SD; n = 10 N = 3). a-b, unpaired, two-tailed Student’s t-test; c, Kruskal-Wallis test; g, One-way ANOVA.
