Figure 2. The effect of FK866 on RAW 264.7 metabolism.

A, B, Glucose consumption and lactate production in the presence and absence of FK866. Cells were incubated in control or FK866 medium for a total of 24 hours. Glucose and lactate was measured in medium supernatants collected over the last 6 hours (18–24 h) of incubation. C, Intracellular ATP levels of RAW 264.7 macrophages treated with 5 nM FK866 for 3, 15, or 24 hours. D, Mitochondrial NAD(P)H-level. Cellular autofluorescence was measured after 24 hour incubation in control medium or medium with 5 nM FK866. E, After 24 hour pre-incubation with or without FK866, oxygen consumption was measured in suspensions of 1×10⁶ cells in either FK866 or control medium. Control and FK866 treated cells were analyzed in parallel on the same day. The basal oxygen consumption was measured where after oligomycin, FCCP, and rotenone was added successively in order to determine the leak respiration, maximal respiration (Max), and residual oxygen consumption (Res). Data in a-c represent means ± SEM of three experiments performed in triplicate, in d means ± SEM of three experiments, and in e means ± SEM of five experiments. (*p<0.05, **p<0.01; unpaired t-test).