Figure 5

Effect of the adenosine kinase inhibitor 5'-ITC on the GAPDH distribution. (A) HDFs were incubated in medium containing 10% FBS and treated with vehicle (DMSO) or various concentrations (0.1-2 µM) of the adenosine kinase inhibitor 5'-ITC 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 + 5'-ITC (µM). (B) 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 5'-ITC (0.1-2 µM) for 2 days. ^***P < 0.001, comparison between the FBS control (0) and FBS + 5'-ITC (µM) after SFM-4d. (C) HDFs were incubated in a culture medium containing 10% FBS for 2 days and then treated with 2 mM AICAR in the absence (0) or presence of various concentrations of 5'-ITC (0.1-2 µM) for 2 days. ^***P < 0.001, comparison between 2 mM AICAR only without compound C (0) and 2 mM AICAR + 5'-ITC (µM). (D) HDFs were serum-depleted by incubating in SFM for 4 days (SFM-4d), and then 10% FBS was re-added to serum-depleted cells with 2 mM AICAR in the absence (0) or presence of various concentrations of 5'-ITC (0.1-2 µM) for 2 days. ^***P < 0.001, comparison between the AICAR/10%FBS control (0) and AICAR/10%FBS + 5'-ITC (µM) after SFM-4d.