Prox1 co-localizes with LSD1/NuRD complex components on <i>CYP7A1</i> promoter.

<p>(A) Occupancy of HNF4α, Prox1, and LSD1/NuRD components on <i>CYP7A1</i> promoter in HepG2 cells. Chromatin immunoprecipitation (ChIP) was performed on chromatin fragments prepared from HepG2 cells using specific antibodies as indicated and corresponding normal IgG as non-specific control. (B) Prox1 co-localizes with LSD1 and HDAC2 on <i>CYP7A1</i> promoter in HepG2 cells. Sequential ChIP was performed on chromatin fragments prepared from HepG2 cells using anti-Prox1 antibodies for first round ChIP (A) and antibodies to HNF4α LSD1 and HDAC2 for second round ChIP, respectively. (C) Occupancy of Prox1, LSD1 and HDAC2 on <i>CYP7A1</i> promoter in mouse liver cells. Chromatin fragments prepared from mouse liver cells were subjected to ChIP using antibodies to Prox1, HNF4α LSD1 and HDAC2 as indicated. In panels A-C, corresponding normal mouse or rabbit IgG was used as non-specific background control for each antibody used. Precipitated <i>CYP7A1</i> promoter segments were detected using quantitative real-time PCR and relative chromatin occupancy was calculated as %input as described in Materials and Methods. In panel A, a control region in downstream <i>CYP7A1</i> mRNA coding sequences (CDS) was also quantitated using real-time PCR in parallel as further demonstration of assay specificity. Means and SD from three independent experiments are presented. Statistically significant enrichment by specific antibodies (<i>P<</i>0.05 in student’s <i>t</i> test) were indicated (*).</p>