Figure 6

Effect of the AMPK inhibitor compound C on the GAPDH distribution. (A) HDFs were incubated in medium containing 10% FBS and treated with vehicle (DMSO) or various concentrations of the AMPK inhibitor compound C (1-10 µM) for 2 days. Cells were immunostained and analyzed by confocal laser scanning microscopy, as described in Figure 1. The number of cells with cytosolic GAPDH alone (cytosol) and cells having nuclear GAPDH with or without cytosolic GAPDH (nuclear +/- cytosol) was counted and the percent distributions are plotted as means ± standard deviations. ^***P < 0.001, comparison between the FBS control and FBS + compound C (µM). (B) HDFs were incubated in medium containing 10% FBS and treated with vehicle (DMSO) or 10 µM of compound C for the times indicated (1-4 days). ^**P < 0.01 and ^***P < 0.001, comparison between the FBS control and FBS + 10 µM compound C. (C) HDFs grown in 10% FBS culture medium were serum-depleted by incubating in SFM for 4 days (SFM-4d) without (0) or with various concentrations of compound C (1-10 µM). ^***P < 0.001, comparison between SFM-4d without compound C (0) and SFM-4d + compound C (µM). (D) HDFs were incubated in medium containing 10% FBS for 2 days and then treated with vehicle (PBS) or 2 mM AICAR in the absence (0) or presence of various concentrations of compound C (1-10 µM) for 2 days. ^***P < 0.001, comparsion between 2 mM AICAR only without compound C (0) and 2 mM AICAR + compound C (µM). The results shown are representative of more than three independent experiments.