Figure 4
LTA induces ROS generation and MMP-9 expression via PKCα. a Cells were pretreated with or without GF109203X (GF, 30 μM) or Gö6976 (Gö, 1 μM) for 1 h before exposure to 50 μg/ml LTA for the indicated time intervals. The conditioned media were analyzed by gelatin zymography. b Cells were pretreated without or with Gö (1 μM) for 1 h and then incubated with LTA for the indicated time intervals (upper part) or 10 min (lower part). The membrane and cytosol fractions were prepared and analyzed by Western blotting using anti-PKC-α, Gαs (as a membrane control) or GAPDH antibody. c Cells were pretreated with or without GF or Gö for 1 h before exposure to LTA for 15 min. The Nox activity and ROS generation were analyzed. d Cells were pretreated with or without Gö (1 μM) for 1 h before exposure to LTA for the indicated time intervals. e Cells were transfected with scramble (scra) or PKCα 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. The cell lysates were analyzed by Western blotting using anti-phospho-serine (p-Ser), p47phox or GAPDH antibody. Data are expressed as the mean ± SEM of three independent experiments. * P < 0.01; # P < 0.01, as compared with the cells stimulated with LTA (c, d) alone. The figure represents one of three similar experiments.