Figure 3

A PFKFB3 small molecule antagonist causes cell death and a reduction of proliferation caused by anti-CD3/anti-CD28-conjugated microbeads after 24–72 hours. Either vehicle (DMSO) or 1–10 μM 3PO was added to human CD3⁺ T cells stimulated with anti-CD3/anti-CD28-conjugated microbeads and IL-2 for 5, 10, 24, 48 and 72 hours and dead (A) and viable (B) T cells were enumerated with a New Brunswick Nucleocounter based propidium iodide staining with and without lysis. (C), Lactate secretion after addition of anti-CD3/anti-CD28-conjugated microbeads in the presence or absence of the indicated concentrations of 3PO for 5–72 hours. (D), Lactate secretion after addition of anti-CD3/anti-CD28-conjugated microbeads in the presence or absence of the indicated concentrations of 3PO for 5–72 hours was normalized to viable T cell counts. Data are representative of three independent experiments. *p < 0.05 (relative to vehicle control).