Figure 3

Involvement of p47 ^phox translocation in LTA-induced MMP-9 expression. Cells were pretreated without or with Apo (10 μM) for 1 h and then stimulated with 50 μg/ml LTA for the indicated time intervals or 10 min. a The whole cell lysates and (b) the membrane fraction were prepared and analyzed by Western blotting using an anti-phospho-serine (p-Ser), p47^phox or Gαs (as a membrane control) antibody. The p47^phox translocation was also observed by immunofluorescent staining. c Cells were transfected with scramble (scra) or p47^phox siRNA for 24 h, followed by stimulation with LTA for 16 h. The conditioned media and cell lysates were analyzed by gelatin zymography or Western blotting analysis of p47^phox and GAPDH. Data are expressed as the mean of three independent experiments. ^* P < 0.05; ^# P < 0.01, as compared with the cells stimulated with vehicle (a) and LTA (a, b) alone. ^§ P < 0.01, as compared with the cells stimulated with LTA (a) alone. The figure represents one of three similar experiments.