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. 2021 Jan 1;11(4):1780–1794. doi: 10.7150/thno.51931

Figure 4.

Figure 4

Antisense eRNA's function in cis relies on new two-looping interaction. (A) Virtual circularized chromosome conformation capture (4C)-seq data from http://promoter.bx.psu.edu/hi-c/chiapet.php showing potential signal binding regions with PSA and FKBP5 promoter in NHEK GM12878 and LNCaP cells. (B and C) The diagram showed that chromosome conformation capture (3C) assay with NcoI digesting in PSA locus and KpnI digesting in FKBP5 locus. Evaluation of the enhancer-promoter-antisense enhancer (gene-ending region) interaction at FKBP5 and PSA target gene loci were showed by chromosome conformation capture (3C) assays. Input meant that the genomic DNA was not crossed link with protein, then was digested by NcoI (for PSA) or KpnI (for FKBP5) and ligated randomly. Input DNA meant genomic random ligated DNA. Error bars represented mean ± SD for triplicate experiments (n=3). P values were shown in the figures. NS, no significance. (D) Evaluation of the enhancer-promoter-antisense enhancer (gene-ending region) interaction at PSA target gene loci by 3C assays in sense eRNA (S-eRNA) or antisense eRNA (As-eRNA) knocked down by LNAs. Error bars represented mean ± SD for triplicate experiments (n=3). P values were shown in the figures. NS, no significance.