Fig 1: Lactate Reduces the Motility of CD4+ T Cells via Reduced Glycolysis and Enhanced FAS(A) Representative western blots (left) and densitometric quantification (right; n = 3) of HK1, HK2, PFK, enolase 1a, and PKM1/2 expression by activated CD4+ T cells treated with sodium lactate (10 mM), or left untreated. Untreated CD4+ T cells (CN, dotted line) set to 1.(B) Representative western blots (left) and densitometric quantification (right; n = 3) of HK1, HK2, enolase 1a, PKM1/2, GCK, and aldolase expression by activated CD4+ T cells treated with sodium lactate (10 mM) and/or SLC5A12 Ab, or left untreated. Untreated CD4+ T cells (CN, dotted line) set to 1.(C) Representative western blots showing mitochondrial and cytosolic HK2 in activated CD4+ T cells treated with sodium lactate (10 mM) for the indicated time points or left untreated (CN). VDAC and ß-actin were used as controls for mitochondrial and cytosolic fraction, respectively. Data representative of n = 2 independent experiments.(D) Representative immunofluorescence images of untreated and lactate-treated CD4+ T cells. Co-staining for hexokinase 2 (green), MitoTracker (red), and DAPI (blue). Scale bar, 10 µm.(E) NADP+ and NADPH intracellular levels in CD4+ T cells (n = 5) treated with sodium lactate (10 mM) for the indicated time points after 72-h activation and shown as NADP+/NADPH ratio. Lactate-untreated CD4+ T cells (CN, dotted line) set to 1.(F) In vitro chemokinesis of activated CD4+ T cells in response to CCL20 (500 ng/mL; n = 4) or CXCL10 (300 ng/mL; n = 3) in the presence of sodium lactate (10 mM) with or without the metabolic drugs C75 (10 µM), TOFA (20 µM), and DHEA (20 µM). Untreated CD4+ T cells (w/o CXCL10, dotted line) were set to 100.(G) In vitro chemokinesis of activated CD4+ T cells (n = 4) from Slc5a12 WT or KO mice in response to CXCL10 (300 ng/mL; 4 h) in the presence of sodium lactate (10 mM). Untreated CD4+ T cells (CN, dotted line) were set to 100.Two-tailed Student’s t test (A), (B), and (E) or one-way ANOVA (F) and (G). Data expressed as mean ± SEM. *p = 0.05; **p = 0.01; ***p = 0.001; #p = 0.05 versus lactate + chemokine. See also Figure S6.