Fig. 5: Regulation of Glut2 expression by HIF-1α and galectin-9.
From: The glucose transporter 2 regulates CD8+ T cell function via environment sensing
a,b, Mouse naive CD8+ T cells were activated by antibodies in the presence or absence of the HIF-1α selective inhibitor PX-478 (20 µM) before analysing surface expression of Glut2 (a) and Glut1 (b). Histograms and mean data ± s.d. from a representative experiment (n = 3, N = 3) are shown. c, C57BL/6J mice received an IP injection of PX-478 (20 mg per kg body weight) for 3 d. Representative histograms and mean data (±s.d.) of Glut2 expression by CD8+ T cells are shown (n = 3, N = 2). d, Glut2 expression by mouse HIF+ or HIF+ CD44hi CD8+ T cells. Representative histograms and mean data (±s.d.) are shown (n = 3, N = 3). e–g, Transcription of Glut1 (e), Glut2 (f) and Gal-9 (g) by mouse HIF+ or HIF− CD44hi CD8+ T cells measured by RT–PCR. Gene expression was normalized to the housekeeping gene tubulin. Control HIF− was set as 1. Error bars show the s.d. (n = 3, N = 3). h, Surface expression of Gal-9 by mouse HIF+ and HIF+ CD8+ T cells. Histograms and mean data (±s.d.) from a representative experiment are shown (n = 9–10, N = 2). i, Mouse naive CD8+ T cells were activated by antibodies for 2 d and subsequently maintained in high (20%) or low (5%) oxygen incubators for a further 24 h. Gal-9 surface expression was then analysed. Histograms and mean data (±s.d.) from a representative experiment are shown (n = 3, N = 2). j, Glut2 expression by mouse Gal-9+ or Gal-9− CD44hi T cells. Histograms and mean data (±s.d.) from a representative experiment are shown (n = 6, N = 2). k, Mouse naive T cells were activated by antibodies 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. Histograms and mean data (±s.d.) from a representative experiment (n = 3, N = 2). l,m, ECAR and OCR by antibody-activated mouse Gal-9+ and Gal-9−CD8+ T cells. l,m, Mean glycolysis and glycolytic capacity (±s.d.; n = 5, N = 2) (l) and mean maximal respiration and spare respiratory capacity (m) are shown (±s.d.; n = 5, N = 2). n,o, Gal-9+ and Gal-9− mice received skin grafts from BL/6.Kd donors (n). CD8+ T cells were depleted by IP injection of 200 μg anti-CD8 at days –1 and +1 (o) (n = 7–8, N = 1). a–d,e–j,l,m, unpaired two-tailed Student’s t-test; k, one-way ANOVA; n,o, log-rank (Mantel–Cox) test.
