Involvement of p47
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), p47phox or Gαs (as a membrane control) antibody. The p47phox translocation was also observed by immunofluorescent staining. c Cells were transfected with scramble (scra) or p47phox 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 p47phox 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.