Skip to main content
. 2008 Dec 12;283(50):35140–35153. doi: 10.1074/jbc.M804540200

FIGURE 5.

FIGURE 5.

Ubiquitinated YY1 physically interacts with HDAC3 in the nuclei of rat hippocampal cells; however, the interaction is markedly reduced after neuronal depolarization. A, HDAC1, HDAC2, and HDAC3 are expressed in cell nuclei of the rat hippocampal cells, and their expression is modified by PTZ treatment. Equal amounts (20 μg) of nuclear cell lysates obtained from the unstimulated (-PTZ) as well as from the PTZ-stimulated (2 h; +PTZ) rat hippocampi were analyzed by Western blot with antibodies against anti-HDAC1, -2, and -3. As a loading control, results of Western blot analysis for histone H3 are shown. B, YY1 physically interacts with HDACs in the nuclei of the rat hippocampal cells. Equal amounts (300 μg) of nuclear cell lysates obtained from the unstimulated (-PTZ) as well as from the PTZ-stimulated (2 h; +PTZ) rat hippocampi were immunoprecipitated (IPα) with anti-HDAC1, anti-HDAC2, or anti-HDAC3 antibody and analyzed for YY1 expression by Western blot. As a positive control, protein nuclear cell lysate obtained from the unstimulated hippocampi has been used (resolved in the input lane). The unmodified YY1 and ubiquitinated YY1 (uYY1) are indicated. C, after PTZ-evoked neuronal activation, an expression of the ubiquitinated form of YY1 is diminished in cell nuclei of the rat hippocampus. Equal amounts of nuclear cell lysates obtained from the unstimulated (-PTZ) as well as from the PTZ-stimulated (2 h; +PTZ) rat hippocampi were immunoprecipitated with anti-YY1 or control (isotype) antibody and analyzed for a ubiquitin expression by Western blot. As a positive control, protein nuclear cell lysates obtained from the unstimulated hippocampi have been used. Please note that the intensity of IgG bands reflects similar loading of immunoprecipitated samples. The ubiquitinated YY1 (uYY1) and IgG heavy chain bands are indicated. D, interaction of HDAC3 with ubiquitinated YY1 is neuronal activity-dependent in cell nuclei of the rat hippocampus. Equal amounts (300 μg) of nuclear cell lysates obtained from the unstimulated (-PTZ) as well as from the PTZ-stimulated (2 h; +PTZ) rat hippocampi were immunoprecipitated with anti-HDAC3 or control (isotype) antibody and analyzed by Western blot for ubiquitin. As a positive control, 20% of protein nuclear cell lysate immunoprecipitated with anti-YY1 antibody from the unstimulated hippocampi has been used. The ubiquitinated YY1 (uYY1) is indicated. In the input lane, 30 μg of hippocampal nuclear proteins has been resolved.