FIG. 4.

Analysis of PML and HDAC interaction in vitro by GST pull-down assay. (A) In vitro-translated ³⁵S-labeled PML protein (IVT PML) was incubated with GST, GST-HDAC1, GST-HDAC2, and GST-HDAC3 immobilized to the glutathione agarose bead. In this assay, GST was used as a negative control. The GST pull-down assay was repeated at least two times, and a weak interaction between HDAC3 and PML was detected consistently. (B) GST-PML pull-down assay of the in vitro-translated ³⁵S-labeled HDAC1 protein (IVT HDAC1). The experiment was performed as described above. GST was used as a negative control in this study. (C) GST-HDAC pull-down assay for PML-RARα and RARα. In this assay, the in vitro-translated and ³⁵S-labeled PML, PML-RARα, and RARα were used as described above. GST-HDAC1 was unable to pull down the PML-RARα and RARα proteins.