Figure 5.

KDM2B Targets the PCGF1/PRC1 Variant Complex Leading to PRC2 Recruitment and Formation of a Polycomb Domain and a Novel System to Ablate KDM2B-Mediated Targeting of PCGF1/PRC1 to Chromatin

(A) ChIP analysis for TetR, RING1B, PCGF1, H2AK119ub1, EZH2, and H3K27me3 across the TetO containing locus in lines expressing TetR, TetR-KDM2B, and TetR-KDM2A. ChIP experiments were performed at least in biological duplicate with error bars showing SEM.

(B) Western blot analysis of PRC1 and PRC2 protein levels after knockdown of PCGF1 in the TetR-KDM2B fusion line.

(C) ChIP analysis for TetR, PCGF1, RING1B, and EZH2 following PCGF1 knockdown in the TetR-KDM2B fusion line. ChIP experiments were performed at least in biological duplicate with error bars showing SEM.

(D) A schematic illustrating tamoxifen (OHT)-dependent removal of the ZF-CxxC domain from both KDM2B isoforms.

(E) Western blot analysis of the Kdm2b^fl/fl cell line following an OHT treatment time course. (^∗) is a nonspecific cross reactive band.

(F) A heat map covering a 10 kb region centered over CpG islands showing KDM2B ChIP-seq in the Kdm2b^fl/fl cells prior to (−) and after 72 hr (+) OHT treatment.

(G) ChIP analysis of KDM2B and PCGF1 at gene-associated CpG islands and gene body regions. ChIP experiments were performed in biological triplicate with error bars showing SEM. See also Figures S4 and S5.