FIGURE 2.

Real-time dynamic imaging of the effects of glucose and insulin on the subcellular localization of FoxO1-EGFP. Single primary β cells derived from mouse islets (A and C) or clonal MIN6 cells (E and G) were infected with FoxO1-EGFP-expressing adenovirus and cultured for 48 h in complete medium and then overnight in 3 mm glucose. Cells were then transferred to the heated stage of a Leica SP2 confocal microscope in KRB containing 3 mm glucose. Images were captured every 2 min up to 60 min, either 15 min (A) or immediately before and then after the addition of 16.7 mm (A) or 30 mm (E) glucose, 20 nm (C) or 200 nm insulin in (C, G) as indicated. Traces in B are typical of the single cells seen in A and are presented to distinguish the glucose-dependent effects above basal fluctuations. All other traces (D, F, H) show the mean ± S.E. from ≥3 independent experiments. Error bars are included only for the 15, 30, and 60 min time points for clarity. *, p < 0.05. Scale bar, 10 μm (5 μm for A).