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. 2017 Mar 30;7:45623. doi: 10.1038/srep45623

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Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Histograms showing caspase 3/7 activity in NP cells derived from (A) rat injured IVD (*p = 0.008), (B,C) human degenerative disc NP treated with pro-inflammatory cytokines, IL-1β and TNFα alone or in presence of CTGF, WISP2 and TGFβ1 (*p < 0.05). Histograms showing caspase 9 activity in NP cells (D) rat injured IVD, (E,F) human degenerative disc NP cells treated with pro-inflammatory cytokines, IL-1β and TNFα alone or in presence of CTGF, WISP2 and TGFβ1(*p < 0.05, **p < 0.005). Each bar in the caspase assays is showing mean ± S.D. of 2 independent experiments done in quadruplets (n = 8). The p-values were determined using paired Student’s t-test. For the combination of IL-1β and TNFα are with respect to no treatment control (NTC), while p-values for the groups containing growth factors (CTGF, WISP2 or TGFβ1) are with respect to the group containing combination of IL-1β and TNFα only. Representative Western blot panels showing expression of MMP-3, MMP-13 and Cox2 in rat healthy IVD NP cells treated with (G) IL-1β alone, (H) IL-1β and TNFα in combination, and in the presence of CTGF, WISP2 or TGFβ1. All Western blots were run under the same experimental conditions as described in Materials and Methods. Histograms showing decreased expression of (I) Cox2, (J) MMP-13 mRNA levels in human degenerative disc NP cells treated with IL-1β and TNFα in presence of CTGF and TGFβ1 in comparison to IL-1β and TNFα only treatments. Each bar in the histogram represents mean ± S.D. of 3 independent experiments done in duplicates (n = 6, *p < 0.001).