Figure 2.
UV or HMBA treatment induces the release of Brd4 from chromatin. (A) LSF (left) and HSF (right) prepared from the UV- or HMBA-treated HeLa cells were subjected to western blotting (WB) for the indicated proteins. (B) Salt-titrated extraction assay of HeLa cells treated with HMBA or UV. The cells were extracted three times with low-salt buffer, and the nuclei were aliquoted and each was extracted with buffer containing indicated concentrations of NaCl. The nuclear pellets were lysed in SDS-loading buffer and analyzed by WB to determine the amount of Brd4 remained in chromatin. (C) The levels of released Brd4 in LSF (top) and the total amount of Brd4 in the cells (bottom) were determined by WB for HeLa cells that were treated with 10 mM HMBA for 4 h, and recovered in normal culture medium for the indicated period.