Figure 7

RAGE-independent NF-κB activation upon S100A4-stimulation of II-11b cells. A. Western blot showing reduced RAGE protein expression by siRNA transfection. 48 hours after transfection the cells were stimulated with S100A4 for one hour and cell lysates subjected to immunoblotting using anti-phospho-IκBα. The results shown are representative of three independent experiments. siNC = siRNA negative control. B. Western blot showing RAGE expression in a panel of cell lines. NF-κB activation refers to the levels of S100A4-induced NF-κB activation demonstrated in a previous study [11], except for osteoblasts, where it refers to IκBα phosphorylation levels shown in Fig. 7C. C. Western blot showing IκBα phosphorylation in human osteoblasts after S100A4 stimulation for the indicated time periods. α-tubulin was used as loading control.