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. 2012 Dec 10;288(5):3174–3183. doi: 10.1074/jbc.M112.414839

FIGURE 3.

FIGURE 3.

PHF6 binds to rDNA promoter and suppresses rRNA synthesis. A, PHF6 binds to rDNA promoter. HeLa cells stably expressing FLAG-tagged PHF6 were generated. ChIP assays were performed by using control IgG or anti-FLAG antibody. Purified immuno-enriched samples were analyzed by real-time PCR using primer pairs that amplify the indicated regions of human rDNA. Error bars, S.D. from at least three independent experiments. B, PHF6 suppresses rRNA synthesis. Total RNA was extracted from the indicated cells, and RT-PCR was performed using the indicated sequence-specific PCRs. C, relative levels of pre-rRNA were determined by real-time quantitative PCR in the indicated cells. Values are obtained by normalizing the rRNA levels with β-actin mRNA levels. Error bars, S.D. (n = 3). D, ribosomal RNA synthesis was assessed by FUrd (10 μg/ml) incorporation in HeLa cells. Cells were labeled with fluorouridine for 10 min and then fixed and immunostained using anti-BrdU antibody. The FUrd incorporation was quantitated. Error bars, S.D. (n = 3).