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. 2018 Apr 24;99(4):749–760. doi: 10.1093/biolre/ioy099

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© The Author(s) 2018. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — NR1H receptors and cell tolerance. Endometrial stromal cells were transfected with scramble siRNA or siRNA targeting NR1H2 or NR1H3 for 48 h. The mRNA expression of each cognate gene was measured by qPCR and normalized to two reference genes, ACTB and RPL19 (A, B). Data are presented as mean (SEM); n = 3 animals. Data were analyzed by Student t-test; values differ from scramble, *** P < 0.001. (C) Endometrial stromal cells cultured with vehicle or transfected with scrambled siRNA, or siRNA targeting NR1H3 (siNR1H3) or NR1H2 (siNR1H2), were analyzed by western blot for ABCA1 and ACTB abundance. (D) Densitometry data for ABCA1 were normalized to ACTB, and are presented as mean (SEM) from three independent experiments. Data were analyzed by ANOVA and Dunnett pairwise multiple comparison t-test; values differ from scramble, * P < 0.05. (E, F) Endometrial stromal cells were transfected with scramble siRNA or siRNA targeting NR1H2 or NR1H3, incubated for 24 h in serum-free medium, and then challenged with control medium black bars () or 100 HU/ml pyolysin red bars () for 2 h. Cell viability was quantified by MTT assay (E), and supernatants were collected to measure leakage of LDH (F). Data are presented as mean (SEM); n = 5 animals. Data were analyzed by one-way ANOVA and Dunnett multiple comparison post-hoc test; values differ from scramble, *** P < 0.001. (Please see the online version for the color figure.)

Inline graphic — NR1H receptors and cell tolerance. Endometrial stromal cells were transfected with scramble siRNA or siRNA targeting NR1H2 or NR1H3 for 48 h. The mRNA expression of each cognate gene was measured by qPCR and normalized to two reference genes, ACTB and RPL19 (A, B). Data are presented as mean (SEM); n = 3 animals. Data were analyzed by Student t-test; values differ from scramble, *** P < 0.001. (C) Endometrial stromal cells cultured with vehicle or transfected with scrambled siRNA, or siRNA targeting NR1H3 (siNR1H3) or NR1H2 (siNR1H2), were analyzed by western blot for ABCA1 and ACTB abundance. (D) Densitometry data for ABCA1 were normalized to ACTB, and are presented as mean (SEM) from three independent experiments. Data were analyzed by ANOVA and Dunnett pairwise multiple comparison t-test; values differ from scramble, * P < 0.05. (E, F) Endometrial stromal cells were transfected with scramble siRNA or siRNA targeting NR1H2 or NR1H3, incubated for 24 h in serum-free medium, and then challenged with control medium black bars () or 100 HU/ml pyolysin red bars () for 2 h. Cell viability was quantified by MTT assay (E), and supernatants were collected to measure leakage of LDH (F). Data are presented as mean (SEM); n = 5 animals. Data were analyzed by one-way ANOVA and Dunnett multiple comparison post-hoc test; values differ from scramble, *** P < 0.001. (Please see the online version for the color figure.)