Figure 2.

Treatment of TNF promoted bone formation of enthesis. (A) Experimental design for inducing bone formation in primary enthesis cells. Both control- and AS-enthesis cells were followed by bone formation inducing conditional medium for the indicated days. As indicated days, the differentiated cells were subjected to various experimental bone forming activities of (B) ALP and Collagen (COL) staining using Sirius red (left) and intracellular ALP activity (right), (C) Alizarin Red S (ARS), Von Kossa (VON), and Hydroxyapatite (HA) stain images, and (D) Quantification of bone formation indicators stain; ARS, VON, and HA (control=4, AS=3). All of the staining images is representative of three independent experiments. (E) RT-qPCR and immunoblotting results of differentiated enthesis cells at 14 days (control=3, AS=3). Each dots display individual values. TNF: tumor necrosis factor, AS: ankylosing spondylitis, ALP: alkaline phosphatase.