Journal: bioRxiv
Article Title: An ancient fecundability-associated polymorphism switches a repressor into an enhancer of endometrial TAP2 Expression
doi: 10.1101/058388
Figure Lengend Snippet: ( A ) Location of rs2071473 with respect to histone modifications that characterize promoters (H3K4me4 ChIP-Seq), enhancers (H3K27ac ChIP-Seq), open chromatin (DNaseI-Seq, FAIRE-Seq), as well as PGR and NR2F2 ChIP-Seq binding sites. Inset, the rs2071473-C allele is predicted to disrupt a DDIT3 binding site. The 1kb region shown in light brown was cloned into the pGL3Basic luciferase reporter vector for functional characterization. ( B ) Luciferase assay results testing the regulatory potential of the T (pGL3Basic-rs2071473C) and C (pGL3Basic-rs2071473T) alleles in endometrial stromal fibroblasts treated with control media (ESFs) or differentiated into DSCs with cAMP/MPA for 48 hours. Data are shown as luciferase expression from the pGL3Basic-rs2071473C or pGL3Basic-rs2071473T reporter relative to renilla expression (pRL-null and empty vector (pGL3Basic) controls. +DDIT3, Relative luciferase expression from the pGL3Basic-rs2071473C or pGL3Basic-rs2071473T in ESFs and DSCs co-transfected with DDIT3. +PGR, Relative luciferase expression from the pGL3Basic-rs2071473C or pGL3Basic-rs2071473T in ESFs and DSCs co-transfected with PGR. +DDIT3 & PGR, Relative luciferase expression from the pGL3Basic-rs2071473C or pGL3Basic-rs2071473T in ESFs and DSCs co-transfected with DDIT3 and PGR. ( C ) DeepSea predicted effects (log 2 fold change) of rs2071473 on regulatory features across cell-types.
Article Snippet: DDIT3 and PGR expression plasmids were also obtained from Genscript.
Techniques: ChIP-sequencing, Binding Assay, Clone Assay, Luciferase, Plasmid Preparation, Functional Assay, Control, Expressing, Transfection