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. 2023 Feb 27;14:1136067. doi: 10.3389/fendo.2023.1136067

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Copyright © 2023 Shi, Zhang, Bao, Liu, Yang and Yin

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The target protein of eugenol in TNBC cells. (A) Work flow of drug affinity responsive target stability experiment; (B) MDA-MB-231 cells were treated with eugenol (20 μM), and lysates were subject to thermolysin digestion and Coomassie Brilliant Blue staining; (C) Enrichment of proteins in the protected band revealed by mass spectrometry analysis; (D) The DARTS assay for target validation. NF-κB protein stability was increased upon eugenol (20 μM) treatment in MDA-MB-231 lysates. Pronase treatment was conducted for 10, 30, and 60 min; (E) The DARTS assay demonstrated the dose-dependent binding of eugenol to NF-κB. Treatment with pronase (5 μg/mL) performed for 30 min “-” represent without the interference of pronase. “+” represent the interference of pronase.