Fig 6.

Colocalization and functional interrelation between endogenous TFAP2C, Myc, and KDM5B at the CDKN1A locus. (A) shTFAP2C-MCF-7 cells were transfected with nonsilencing control siRNA (nsRNAi) or siRNA against Myc (siMyc) or KDM5B (siKDM5B) or treated with doxycycline (Dox) to induce shTFAP2C and harvested 72 h later for use in ChIP with antibodies against IgG (control), TFAP2C, (KDM5B, or Myc. Precipitates were analyzed by qPCR using primer pairs specific to 3 distinct regions (−2290/−2185, −935/−864, and −21/+44) across the CDKN1A 5′ sequence, as indicated by the double-headed arrows. (B) ChIP was performed as described for panel A using antibodies specific for histone 3 and H3K4-me3. qPCRs were performed using the −21/+44 CDKN1A proximal promoter-specific primers. Results are shown as fold enrichment above the IgG (background) control (A) or as an H3K4me3/H3 ratio (B). Data were averaged from three independent experiments, ± standard errors.