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Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure
Examination of transcriptional regulation occurring during pregnancy establishment and maintenance requires the identification of endogenous reference genes characterized by high expression stability. Since the expression of some reference genes may be modulated by pregnancy-associated hormones, the...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9325760/ https://www.ncbi.nlm.nih.gov/pubmed/35882953 http://dx.doi.org/10.1038/s41598-022-17069-3 |
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author | Sozoniuk, Magdalena Jamioł, Monika Kankofer, Marta Kowalczyk, Krzysztof |
author_facet | Sozoniuk, Magdalena Jamioł, Monika Kankofer, Marta Kowalczyk, Krzysztof |
author_sort | Sozoniuk, Magdalena |
collection | PubMed |
description | Examination of transcriptional regulation occurring during pregnancy establishment and maintenance requires the identification of endogenous reference genes characterized by high expression stability. Since the expression of some reference genes may be modulated by pregnancy-associated hormones, the goal of our study was to identify suitable reference genes unaffected by hormonal treatment. In our study bovine caruncular epithelial cells were subjected to progesterone, estrogen and prostaglandin F(2α) treatment. Ten candidate reference genes (ACTR1A, CNOT11, HDAC1, HPRT1, RPL19, RPS9, SDHA, SUZ12, UXT and ZNF131) were evaluated with the use of four approaches (geNorm, NormFinder, BestKeeper, delta Ct). We found that RPS9 and SUZ12 displayed the highest expression stability in the tested material. Moreover, HPRT1 and SDHA were found inappropriate for RT-qPCR data normalization as they demonstrated the highest expression variability out of all candidates analysed. Hence geNorm calculations shown that the use of just two best-performing genes would be sufficient for obtaining reliable results, we propose that RPS9 and SUZ12 be used as suitable endogenous controls in future studies investigating gene expression in normal and compromised pregnancies. |
format | Online Article Text |
id | pubmed-9325760 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-93257602022-07-28 Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure Sozoniuk, Magdalena Jamioł, Monika Kankofer, Marta Kowalczyk, Krzysztof Sci Rep Article Examination of transcriptional regulation occurring during pregnancy establishment and maintenance requires the identification of endogenous reference genes characterized by high expression stability. Since the expression of some reference genes may be modulated by pregnancy-associated hormones, the goal of our study was to identify suitable reference genes unaffected by hormonal treatment. In our study bovine caruncular epithelial cells were subjected to progesterone, estrogen and prostaglandin F(2α) treatment. Ten candidate reference genes (ACTR1A, CNOT11, HDAC1, HPRT1, RPL19, RPS9, SDHA, SUZ12, UXT and ZNF131) were evaluated with the use of four approaches (geNorm, NormFinder, BestKeeper, delta Ct). We found that RPS9 and SUZ12 displayed the highest expression stability in the tested material. Moreover, HPRT1 and SDHA were found inappropriate for RT-qPCR data normalization as they demonstrated the highest expression variability out of all candidates analysed. Hence geNorm calculations shown that the use of just two best-performing genes would be sufficient for obtaining reliable results, we propose that RPS9 and SUZ12 be used as suitable endogenous controls in future studies investigating gene expression in normal and compromised pregnancies. Nature Publishing Group UK 2022-07-26 /pmc/articles/PMC9325760/ /pubmed/35882953 http://dx.doi.org/10.1038/s41598-022-17069-3 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Sozoniuk, Magdalena Jamioł, Monika Kankofer, Marta Kowalczyk, Krzysztof Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
title | Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
title_full | Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
title_fullStr | Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
title_full_unstemmed | Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
title_short | Reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
title_sort | reference gene selection in bovine caruncular epithelial cells under pregnancy-associated hormones exposure |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9325760/ https://www.ncbi.nlm.nih.gov/pubmed/35882953 http://dx.doi.org/10.1038/s41598-022-17069-3 |
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