Figure 1

Activation of human CD3⁺T cells increases PFKFB3 expression and intracellular F2,6BP. CD3⁺ T cells were isolated by negative selection and then plated at a final concentration of 1 x 10⁶ cells/ml in the absence or presence of anti-CD3/anti-CD28-conjugated microbeads for 5, 10 and 24 hours. (A), PFKFB1-4 and TIGAR mRNA expression was determined using real-time RT-PCR analyses. (B), PFKFB2-4, TIGAR, CD69 and β-actin protein expression was determined by Western blot analyses. (C), Intracellular F2,6BP concentration was determined using a coupled enzyme assay. (D), Densitometric analysis of PFKFB2-4, TIGAR, CD69 and β-actin protein expression. Data are representative of three independent experiments. *p < 0.05 (relative to expression or concentration at 0 hours).