Figure 4

Effect of AICAR on GAPDH distribution. (A) HDFs were incubated in medium containing 10% FBS and treated with vehicle (PBS) or various concentrations of AICAR (0.5-2 mM) for 4 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 + AICAR (mM). (B) HDFs were incubated in medium containing 10% FBS and treated with vehicle (PBS) or 2 mM AICAR for the times indicated (1-4 days). ^***P < 0.001, comparison between the FBS control and FBS + 2 mM AICAR. (C) HDFs were serum-depleted by incubating in SFM for 4 days (SFM-4d), and then 10% FBS was re-added to serum-depleted cells and treated with vehicle (0) or various concentrations of AICAR (0.5-2 mM) for 2 days. ^***P < 0.001, comparison between the FBS control (0) and FBS + AICAR (mM) after SFM-4d. (D) HDFs were serum-depleted by incubating in SFM for 4 days, and then either 10% FBS, 100 ng/ml PDGF, or 30 µM LPA was re-added to serum-depleted cells in the absence (-) or presence (+) of 1 mM AICAR for 2 days. ^**P < 0.01 and ^***P < 0.001, comparison between the absence (-) and presence (+) of 1 mM AICAR. The results shown are representative of more than three independent experiments.