Journal: Osteoarthritis and cartilage

Article Title: Age and oxidative stress regulate Nrf2 homeostasis in human articular chondrocytes

doi: 10.1016/j.joca.2023.05.004

Figure Lengend Snippet: (A) siRNA mediated knockdown of Nrf2 in normal chondrocytes (ages 32 to 45, n=4) was performed by nucleofection yielding an ~89% depletion of Nrf2 compared to control siRNA. Nrf2 and its target protein levels were measured by immunoblotting and β-tubulin served as the loading control. The band intensity was normalized to β-tubulin. Results shown are mean ± SD. (B) siRNA knockdown of Nrf2 was performed in normal chondrocytes (ages 42 to 55, n=4) and then the cells were treated with RTA-408 to test for the specificity of Nrf2 activation. After nucleofection, the transduced cells were treated with RTA-408 for 48 hours. Repeated measures ANOVA was used for statistical analysis.

Article Snippet: Cells were harvested for nucleofection using collagnease and pronase, centrifuged, and resuspended in Amaxa primary cell nucleofection reagent.

Techniques: Knockdown, Control, Western Blot, Activation Assay