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Ligand-dependent activation of ERs, both ERE and non-ERE-mediated, attracts co-regulator molecules that modify the chromatin state, thereby recruiting or hindering the transcriptional complex and representing another level of control in ER gene regulation [ 61, 63, 79].
The ligand ER complex in the nucleus interacts with both ERE or non-ERE (tethered) sequences (Couse and Korach 1999).
For ERβ ERE-mediated activation, both ERE reporters exhibited responses to E2 in HepG2 cells.
Our analysis revealed that the both ERE sequences are 100% conserved between Rhesus and human, whereas the 3'-ERE sequence also shares 79% identity with mouse, indicating that the 3'-ERE is more evolutionarily conserved.
They included oestrogen regulated genes resulting from interactions at both ERE and AP-1 sites.
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Disruption of both EREs abrogated ER binding and nearly abolished p53 binding to the p53 RE-T region, consistent with the results of the functional assays.
Given that MCF-7 is an ER-positive breast cancer cell line, HRG-induced transcription may involve both ERE- and non-ERE-dependent regulation.
ER-dependent regulation of PR is reported to be mediated by both EREs and a Sp1 site (Chauchereau et al., 1991; Petz et al., 2004).
EMSA analysis using nuclear extracts of E2-treated cells and natural sequences, including these putative EREs, indicated that ERβ – the only isoform expressed in U-937 cells – specifically recognized both EREs because ERβ DNA complexes were efficiently competed by the corresponding unlabelled probe and supershifted by the anti-human ERβ (L-20) antibody.
Gene reporter assays clearly showed that in MCF7 cells both EREs needed to be inactivated to impair p53-dependent transactivation (Figure 1A).
Here we report the identification of a second ER ½ site (ERE2) located 145 bp downstream of the RE-T and establish that both EREs can impact p53-mediated transactivation of FLT1-T in a manner that is cell type and ER level dependent.
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