Epigenome and transcriptome profile of the individual myometrium biopsy. Active genes are defined as FPKM ≥ 1. N.D., not determined.

Putative enhancers in term pregnant human myometrial tissues. (A) Distinct and common putative enhancers in term pregnant myometrial biopsies from three subjects. Genomic regions with H3K27ac and H3K4me1 double positive histone marks are defined as putative active enhancers. (B) Association of commonly shared putative enhancers with active genes. The association between an interval and an active gene is defined by locating within 100 kb vicinity of each other. (C) Over-Represented transcription factor binding motifs in putative enhancers. A subset of enriched motifs that are relevant to myometrial homeostasis in the 13090 H3K4me1/H3K27ac-positive putative enhancer regions is displayed.

Putative super enhancers in term pregnant human myometrial tissues. (A) Number of super enhancers mapped in tissues of each individual human subject. (B) Association of commonly shared super enhancers with active genes in human myometrium. (C) Enriched gene ontology terms in the 355 active genes that are associated with super enhancers.

Identification of enhancers for the PLCL2 gene. (A) UCSC Genome Browser track view of the human PLCL2 locus marked with gRNA targeting locations. (B-C) Relative PLCL2 mRNA levels measured by qRT-PCR in hTERT-HM cells (B) or T-HESC cells (C) that express denoted gRNAs with the CRISPR activator (N=3 with technical duplicates). ****, p < 0.0001; ***, p < 0.001; **, p < 0.01; *, p < 0.05 by Unpaired t-test.

Candidate upstream regulators that mediate PLCL2-5 enhancer’s regulatory effect on PLCL2 expression.

PGR regulation of PLCL2 expression. (A) PGR mRNA abundance in hTERT-HM cells with the PGR-promoter targeting (PGR640) or non-targeting gRNAs under the CRISPR activation system. (N=3 with technical qPCR duplicates). (B) Protein abundance of in hTERT-HM cells with the PGR-promoter targeting (PGR640 and P6SM) or non-targeting gRNAs under the CRISPR activation system. Protein extracts from unmanipulated MCF7 and HEK293T cells serve as positive and negative control for the PGR presence. (C) PLCL2 mRNA abundance in in hTERT-HM cells with the PGR-promoter targeting (PGR640) or non-targeting gRNAs under the CRISPR activation system. (N=3 with technical qPCR duplicates). (D) Pearson correlation between PLCL2 mRNA levels and inferred PGR activities (T-Scores) in 43 human myometrial specimens. **, p < 0.01; *, p < 0.05 by Mann-Whitney test (A) and Unpaired t-test (C).