FIG. 7.

Lsh dissociates from chromatin after TSA treatment. (A) Lsh associates with DAPI in hypomethylated MEFs. Lsh wild-type and Lsh^−/− MEFs were transfected with a GFP-tagged Lsh expression vector and examined after 24 h. (B) TSA induces hyperacteylation. 3T3 cells were treated in the presence of 75 ng of TSA/ml, and extracts were examined by Western analysis with antibodies raised against histone 3 (H3), acetylated H3 (AcH3), or acetylated H4 (AcH4; equal loading of protein extracts). (C) Lsh dissociates from chromatin after TSA treatment and reassociates after recovery. After TSA treatment or after an additional 24 h of recovery, 3T3 cells were extracted with Triton X, and the different fractions were examined by Western analysis for the presence of induced Lsh; Vimentin served as a control. Triton X-resistant (lanes R) and soluble (Triton flush out [lanes F]) (extracts were prepared from equal number of cells) fractions are shown.