FIG. 3.

siRNA knockdown of BAT3 eliminates BAT3 binding to the promoter regions of BRCA1 and Myc, decreases gene expression, and alters H3K4 dimethylation. (A and B) BAT3 is not required for BORIS DNA binding. HCT116 cells were treated with either nonsense (control) or BAT3 siRNA, and ChIP analysis was done followed by QPCR to with primers to the Myc (A) or BRCA1 (B) promoters (see Fig. S5 in the supplemental material). (C) siRNA knockout of BAT3 decreases BRCA1 and MycR RNA levels. HCT116 cells were treated with nonsense (Cont) or BAT3 siRNA (in duplicate); RNA was isolated; and BRCA1 and Myc expression was determined by RT-PCR that was done in triplicate. Error bars represent one standard deviation about the arithmetic mean, and statistical significance was established by Student's t test. *, P < 0.05. (D) siRNA knockout of BAT3 decreases histone H3K4 dimethylation. HCT116 cells were treated with nonsense (Cont) or BAT3 siRNA and harvested for ChIP using either an H3K4 or H3K9 antibody followed by QPCR with primers to either the Myc (left panel) or BRCA1 (right panel) promoter. All ChIP experiments were done in triplicate, and error bars represent one standard deviation about the arithmetic mean. Statistical significance was established by Student's t test. *, P < 0.05 by t test.