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. 2000 Jun 27;97(14):7835–7840. doi: 10.1073/pnas.140199597

Figure 3.

Figure 3

Phosphorylation-dependent binding of 14-3-3 to HDAC4 and HDAC5. (A) Association of HDAC4 and HDAC5 with 14-3-3 and HDAC3 depends on the phosphorylation state of the proteins. HDAC4-FLAG and HDAC5-FLAG were transiently expressed in TAg Jurkat cells. Forty-eight hours post-transfection, the cells were treated for 1.5 h with staurosporine and calyculin A. The immunopurified complexes were subjected to Western blot analysis and were tested for HDAC activity, as described in Materials and Methods. (B) Association of endogenous HDAC4 with 14-3-3 β in NIH 3T3 cells is phosphorylation-dependent. 14-3-3 β immunoprecipitates from NIH 3T3 cells that had been treated with staurosporine or calyculin A for 1.5 h were analyzed for the presence of endogenous HDAC4 by Western blotting.