FIG. 6.

Effect of trien on cell signaling responses to pioglitazone. A: H4IIE cells were grown in serum-free medium for 18 h in the presence or absence of 10 mM trien, followed by stimulation with or without differing doses of pioglitazone, as indicated. Lysates were prepared for immunoblotting with the antibodies indicated as described in research design and methods. B: H4IIE cells were grown in serum-free medium for 18 h in the absence or presence of 10 mM trien. Individual dishes were then stimulated without or with 15 μM pioglitazone (Pio) for 3 h. Total of 2 mM metformin (Met) was added to further dishes for 3 h as a positive control, and then the samples were prepared for AMPK assay as described in research design and methods. Statistical analysis was carried out as in Fig. 1D. ***P < 0.001.