FIGURE 3.

Influence of transitory knock-down of Ank on Wnt-5a production. A, Western blot analysis of conditioned supernatant. Supernatants were subjected to Western blotting. Quantification of protein content of the loaded volumes of supernatant (in micrograms) was done using the bicinchonic acid method. Images presented are representative of three independent experiments. B, effect of conditioned supernatant on Sox-9 and type II collagen expression. Conditioned supernatants were neutralized or not for 1 h at 37 °C, using either mock (anti-β-actin) or anti-Wnt-5a antibodies. Total RNA was extracted from chondrocytes transfected with either scramble or Ank siRNA, and subjected to RT-qPCR analysis. The levels of mRNA of interest were normalized to that of S29 (used as reference gene). Results are presented in histograms as means (± S.D.) over S29 value (n = 3). Statistically significant differences from the Scramble are indicated as *, p < 0.05, and from mock as #, p < 0.05.