Fig. 4

GLUT1 drives Tp-induced cytokine production and increased glycolysis. A THP-1 cells were treated with PBS, DTp, and Tp (MOI of 2:1) for 24 h. Gene expression of different glucose transporters was measured by qRT-PCR, normalized to β-actin. B The mRNA levels of SLC2 A1 after THP-1 cells were treated with PBS, DTp, and Tp (MOI of 1:1, 2:1) for 24 h. C The expression of GLUT1 protein after THP-1 cells were treated with different concentrations of Tp (MOI of 1:1, 2:1, 4:1) for 24 h was measured by Western blot. Expression of GLUT1 protein after THP-1 cells were treated with Tp (MOI of 2:1) for different times was measured by Western blot. D GLUT1 membrane localization in THP-1 cells treated with PBS or Tp was assessed by immunofluorescence microscopy. Yellow arrows indicate the enlargement of some cells. The imaging is representative of three experiments. F-I THP-1 cells were pretreated with the GLUT1-specific inhibitor BAY876 (25, 50 nM) for 2 h and then cocultured with Tp (MOI of 2:1) for 24 h. The mRNA levels of (F) SLC2 A1, (G) HK1, (H) HK2, and (I) LDHA were detected by qRT-PCR. E Glucose uptake by THP-1 cells was measured by flow cytometry. J The time course of real-time changes in extracellular acidification after the glycolysis assay. K IL-6, (L) IL-8, and (M) CCL2 concentrations in the cell culture supernatants of THP-1 cells were quantified by ELISA. The data are shown as the mean ± SD. Statistical significance tested by one-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, ns: no significant, n = 3