Histone Deacetylase Inhibitors Increase p27^Kip1 by Affecting Its Ubiquitin-Dependent Degradation through Skp2 Downregulation

Figure 2

Effect of HDACIs on the cellular content. (a) On the left, K562 cells were incubated with different HDACIs for 24 and 48 hours at the concentrations showed. Then, total cell extracts were prepared and the content of was determined. On the right, K562 cells were incubated with NAM and BuA at the concentration reported in Figure 1(a) for 24 hours. Then, total cell extracts were prepared and the content of was determined. Actin was also analyzed for confirming equal loading. (b) EPN and K562 cells were incubated for different time periods with 1 mM BuA. Subsequently, and actin contents were determined by immunoblotting. Actin was used for verifying equal loading. (c) K562 cells were incubated for 16 hours with 1 mM BuA. Then, the nuclear and cytosolic compartments were prepared. Finally, the content of and HDAC1 was determined by immunoblotting. Histone H3 and β-tubulin were used as markers of nuclear and cytosolic fraction, respectively, and for equal loading evaluation.

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