Fig 1: Anti-inflammatory action of hsa-miR-125b-5p against IL-1β induced inflammatory signaling in human OA chondrocytes. IL-1β through IL-1R activates TRAF6 mediated signaling to the nuclear transcription factor NF-κB via activation of MARKs induced production of inflammatory mediators including MMP-13 in human OA chondrocytes. MicroRNA-125b-5p regulates MMP-13 expression via TRAF6 3′UTR. Abbreviations: OA: osteoarthritis; IL-1β: interleukin-1 beta; IL-1R, IL-1 receptor; TRAF6: TNF receptor associated factor 6; 3′UTR: three prime untranslated region; MAPK: mitogen activated protein kinase; NF-κB: nuclear transcription factor; IκBα: inhibitor of kappa B protein; MMPs: matrix metalloproteinases.
Fig 2: MicroRNA hsa-miR-125b-5p regulates MMP-13 expression in human OA chondrocytes. (A) MMP-13 gene expression in IL-1β stimulated chondrocytes transfected with miR-125b mimic. @p < 0.01 versus #. (B) MMP-13 activity in the culture supernatant in OA chondrocytes transfected with miR-125b mimic. The production of MMP-13 was determined in cell culture supernatant by gelatin zymography (full length zymographic expression is provided in Supplementary Information File). Equal volumes of culture supernatant were loaded on polyacrylamide gel. The MMP-13 positive control (EMD Chemicals) was also used. Band images were digitally captured and the band intensities (pixels/band) were obtained using the Un-Scan-It software and are expressed in average pixels. Unstimulated chondrocytes were used as controls and expression of RNU6B/GAPDH was used as an endogenous control.
Fig 3: MicroRNA hsa-miR-125b-5p regulates NF-κB signaling events in human OA chondrocytes. (A) Inhibition of NF-κB p65 activity in miR-125b mimic transfected human chondrocytes. @p < 0.01 versus #. (B) Inhibition of NF-κB p50 activity in miR-125b mimic transfected human chondrocytes. @p < 0.01 versus #. (C) Inhibition of IκBα activation in miR-125b mimic transfected human chondrocytes (full length blot is provided in Supplementary Information File). @p < 0.05 versus #. (D–F) Parthenolide (IKK inhibitor) negatively co-regulated hsa-miR-125b-5p expression (D) and MMP-13 mRNA expression (E) and production (F) in IL-1β-stimulated human chondrocytes #p < 0.05 versus parthenolide treated chondrocytes. Unstimulated chondrocytes were used as controls and expression of RNU6B/GAPDH was used as an endogenous control. Band images were digitally captured and the band intensities (pixels/band) were obtained using the Un-Scan-It software and are expressed in average pixels.
Supplier Page from Abcam for Recombinant human MMP13 protein