(A) The truncated ZFP-1 protein similar to that present in the zfp-1(ok554) mutant strain does not interact with DOT-1.1. Top, a schematic of the two FLAG-tagged WT ZFP-1 isoforms and a truncated ZFP-1 protein missing the C-terminal portion, which is identical to that present in the zfp-1(ok554) mutant. Bottom, western blots with an antibody against DOT-1.1 and an antibody against ZFP-1 of total lysates (left) and proteins immunoprecipitating with an antibody against FLAG (right) in indicated transgenic strains or WT nontransgenic worms, rightmost lanes.
(B) Venn diagram showing an overlap between ZFP-1 and DOT-1.1 target genes identified by ChIP-chip. A Fisher’s exact test was used for calculating p values for overlaps.
(C) Example of a genomic region with ZFP-1 (top) and DOT-1.1 (bottom) ChIP-chip binding enrichment at the promoters of target genes (grey bars). Gene models are based on the UCSC Genome Browser (WS220/ce10).
(D) ChIP-qPCR results with antibodies against DOT-1.1 in WT and zfp-1(ok554) demonstrating the role of ZFP-1 in recruiting DOT-1.1 to the promoters of the target genes.
Here, and in the following figures, “P” after the gene name indicates promoter, and “C” after the gene name indicates coding region. The results of three independent experiments are shown. Error bars represent SD.
See also Figures S1–S4.