FIGURE 6.

PHF6 deficiency leads to increased DNA damage at the rDNA locus. A, PHF6 deficiency induced DNA damage. PHF6-depleted HeLa cells or control cells were lysed and subjected to Western blotting (WB) using the indicated antibodies. B, U2OS-DR-GFP cells were transfected with 1 μg of pCBA-I-SceI plasmid. 18 h later, cells were collected, and cell lysates were subjected to ChIP using γ-H2AX antibody. Purified immuno-enriched samples and genomic DNA samples (as input) were used for sequence-specific PCRs. C, PHF6 deficiency induced DNA damage at the rDNA locus. PHF6 depletion or control cells or reconstitution cells were collected and subjected to ChIP by using γ-H2AX antibody. Purified immuno-enriched samples were analyzed by sequence-specific PCRs. The PCR primers target to the rDNA region or β-actin region. D, purified immuno-enriched samples from C were analyzed by real-time PCR using primer pairs that amplify the region of human rDNA. Error bars, S.D. from at least three independent experiments.