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. 2017 Dec 6;7:17050. doi: 10.1038/s41598-017-17138-y

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Mevalonate pathway inhibitors modulate the impact of PLO on cell viability. (A) BESC isolated from 4 animals, or (B) HESC from 3 independent passages were incubated for 48 h with control serum-free medium, or media containing the indicated concentrations of atorvastatin, alendronate or zaragozic acid, and then challenged with control media (■) or media containing PLO () using 100 HU/ml for BESC and 200 HU/ml for HESC, for 2 h. Cell viability was determined by MTT assay. Data are expressed as mean (SEM), and were analyzed by one-way ANOVA with Dunnett’s multiple comparison post hoc test; values differ from PLO treatment with no inhibitor, *P < 0.05, **P < 0.01, ***P < 0.001. In independent experiments, BESC isolated from 4 animals were incubated for (C) 24 h or (D) 48 h in serum-free media containing vehicle, MBCD or 10 μM zaragozic acid. Cells were then challenged with 100 HU/ml PLO, and cell viability was monitored every hour using the Alamar Blue assay. Data are expressed as mean (SEM) percentage viability of control.

Inline graphic — Mevalonate pathway inhibitors modulate the impact of PLO on cell viability. (A) BESC isolated from 4 animals, or (B) HESC from 3 independent passages were incubated for 48 h with control serum-free medium, or media containing the indicated concentrations of atorvastatin, alendronate or zaragozic acid, and then challenged with control media (■) or media containing PLO () using 100 HU/ml for BESC and 200 HU/ml for HESC, for 2 h. Cell viability was determined by MTT assay. Data are expressed as mean (SEM), and were analyzed by one-way ANOVA with Dunnett’s multiple comparison post hoc test; values differ from PLO treatment with no inhibitor, *P < 0.05, **P < 0.01, ***P < 0.001. In independent experiments, BESC isolated from 4 animals were incubated for (C) 24 h or (D) 48 h in serum-free media containing vehicle, MBCD or 10 μM zaragozic acid. Cells were then challenged with 100 HU/ml PLO, and cell viability was monitored every hour using the Alamar Blue assay. Data are expressed as mean (SEM) percentage viability of control.