Immunodepleted CysC or E64 does not stimulate mineralization in bone marrow-derived osteoblastic cells. (A) Changes in cathepsin activity with increased anti-CysC antibody (αCysC). Immunodepletion with anti-CysC antibody (0–7 μM) to examine the inhibitory effect of CysC (10 μM) on the cysteine proteinase activity of mouse
Zanamivir extracts with Z-Phe-Arg-MCA. The data are means ± SD of values from three independent experiments. (B,C) Effects of CysC, iCysC, anti-CysC antibody, or E-64 on calcium and phosphate deposits in de novo nodules after 28 days. (B) Calcium content in the nodules. (C) Total area of von Kossa-positive nodules. ??p
cells (CTRL); white column, CysC-treated cells (CysC); dark-gray column, iCysC-treated cells; light-gray column, anti-CysC antibody-treated cells; checked column, E-64-treated cells (E-64). Each value is mean ± SD of triplicate determinations. Error bars show the SD. (D) Effects of CysC, iCysC, anti-CysC antibody, or E-64 on calcium and phosphate deposits in de novo nodules after 28 days. Upper panel, alizarin red staining; lower panel, von Kossa staining.
