Fig. 5: NIPBL and BRD4 stabilize each other on the chromatin.

a Localization of NIPBL and BRD4 at the promoters of the indicated genes was assessed through ChIP analysis with the indicated antibodies (α-) in comparison with normal rabbit IgG. Relative ChIP levels are represented. b Localization of BRD4∆C at the promoters of the indicated genes was assessed through ChIP analysis with FLAG antibodies in comparison with normal mouse IgG. FLAG signal was determined in cells transfected either with the expression construct for the BRD4∆C protein as with empty vector. Relative ChIP levels are represented. c NIPBL and BRD4 localization at the promoters of the indicated genes was assessed through ChIP analysis after Nipbl or Brd4 knockdown (siNip#1, Nipbl siRNA #1, and siB4, Brd4 siRNA, respectively) in comparison with control conditions (siC, Control siRNA). Relative ChIP levels are represented. d NIPBL and BRD4 localization at the promoters of the indicated genes was assessed through ChIP analysis after transfection of the BRD4∆C expression construct (B4∆C) in comparison with control transfections with empty vector. Relative ChIP levels are represented. e Schematic representation of the endogenous wild-type BRD4 and the expressed truncated BRD4∆C protein, indicating the regions recognized by the antibodies (α-) used for ChIP analysis. f Expression levels of the indicated genes, after transfection of the BRD4∆C expression construct were determined by qPCR. Relative levels of expression are represented. Cells transfected with empty vector were used as control. Values are means ± s.d. from three independent experiments analyzed in triplicate. Statistical significance of differences between the different conditions and IgG (a, b), control siRNA (siC) (c), or empty vector (d, f), is indicated on top of each bar. Statistical significance of differences between other conditions is also indicated with a line. Significance was analyzed by Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001)