Figure 3.

PRC2 Binding and H3K27me3 Deposition at Polycomb Target Sites Rely on PRC1 Catalytic Activity

(A) Genomic snapshot of a Polycomb target gene showing cChIP-seq for RING1B, H2AK119ub1, SUZ12, H3K27me3, JARID2, AEBP2, PCL2, and EPOP in PRC1^CPM cells.

(B) Metaplot analysis of SUZ12 cChIP-seq at PcG-occupied sites in PRC1^CPM and PRC1^CKO cells.

(C) As in (B) for H3K27me3 cChIP-seq.

(D) Heatmap analysis of cChIP-seq data shown in (B) and (C). Genomic regions were sorted based on RING1B occupancy in untreated PRC1^CKO ESCs.

(E) A scatterplot comparing the log2 fold changes in SUZ12 levels at PcG-occupied sites in PRC1^CPM and PRC1^CKO ESCs. R² represents coefficient of determination for linear regression and cor denotes Pearson correlation coefficient.

(F) As in (E) for H3K27me3 cChIP-seq.

(G) Metaplot analysis of PRC2.2-specific subunits JARID2 and AEBP2 cChIP-seq at PcG-occupied sites in PRC1^CPM cells.

(H) As in (G) for PRC2.1-specific subunits PCL2 and EPOP.

(I) Heatmap analysis of cChIP-seq data shown in (G) and (H). Genomic regions were sorted as in (D).

(J) Maximum intensity projections of JF₅₄₉-Halo-SUZ12 signal in PRC1^CPM ESCs. Examples of SUZ12 nuclear foci (Polycomb bodies) are indicated by arrowheads. Scale bar is 5 μm.

(K) Boxplots comparing the number of JF₅₄₉-Halo-SUZ12 nuclear foci in PRC1^CPM ESCs before (n_cells = 55) and after (n_cells = 52) OHT treatment. Cells from two independent experiments were analyzed. P values denote statistical significance calculated by a Student’s t test.

See also Figure S3.