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. 2017 Jun 23;7:4112. doi: 10.1038/s41598-017-04384-3

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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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IL-1β upregulated asporin expression in human nucleus pulposus cells via the NF-κB p65 pathway. (a) Immunofluorescence analysis of p65 in human nucleus pulposus cells induced by 10 ng/ml IL-1β with or without the p65 inhibitor Bay 11. (b) Quantification of asporin mRNA expression in human nucleus pulposus cells treated with 10 ng/ml IL-1β with or without Bay 11. (c) Quantification of asporin secretion in the culture supernatant of human nucleus pulposus cells treated with 10 ng/ml IL-1β with or without Bay 11. (d) Western blot analysis of asporin expression in human nucleus pulposus cells treated with 10 ng/ml IL-1β with or without Bay 11. (e and g) Western blot and real time RT-PCR analysis of asporin expression in human nucleus pulposus cells treated with 10 ng/ml IL-1β in the presence or absence of p65 shRNA. (f–h) Western blot and real time RT-PCR analysis of asporin expression in human nucleus pulposus cells induced by 10 ng/ml IL-1β with or without p65 overexpression. p65 over means p65 overexpression; Scramble and Mock represent the negative controls of p65 shRNA and p65 overexpression. *P < 0.05, **P < 0.01, ***P < 0.001. P-values were analyzed by one-way ANOVA. All data are representative of three independent experiments and are means ± SEM. Uncropped images of the blots for Fig. 3d–f are shown in supplementary Figure 3.

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