Figure 5.

Role of annexin II and NF-κB. (A, a) Prefusion osteoclast incubation with Biotin^hbdPRELP (B^hbdPRELP; 10 min) and immunofluorescence detection of annexin II. (b) Immunofluorescence detection of chondroitin sulfate and annexin II. (B) Vital incubation of prefusion osteoclasts with Biotin^hbdPRELP in the presence of anti-CD44, anti–annexin II antibody, and an irrelevant IgG. Biotin-594 streptavidin (streptav) served as a negative control in the absence of Biotin^hbdPRELP. (C) Prefusion osteoclasts, treated as indicated, were incubated with an anti–p65NF-κB antibody followed by incubation with FITC-conjugated secondary antibody (NF-κB; left) and with propidium iodide (PI; middle) to detect nuclei. (D) Colorimetric p65NF-κB DNA binding assay of prefusion osteoclast lysates treated with vehicle or 15 µM ^hbdPRELP. (E) RAW264.7 cells were treated with control peptide or ^hbdPRELP and subjected to the luciferase assay as described in Materials and methods. (D and E) Results are the mean ± SEM of three independent experiments (*, P < 0.05).