Fig. 5

Effects of rottlerin in PMA-stimulated activation of transcription factors such as AP-1. a Cells were pretreated with tanshinone IIA (TSIIA, 10 μM) and then incubated with PMA (1 μM) for the indicated time intervals. The conditioned media were collected and assayed for MMP-9 expression by gelatin zymography. b, d Cells were pretreated with TSIIA (10 μM), NAC (5 mM), Apo (10 μM), PD98059 (PD, 10 μM), and Rott (1 μM) for 1 h and then stimulation with PMA (1 μM) for 30 min (b) or 16 h (d). The total RNA was collected and analyzed c-fos mRNA (30 min) and MMP-9 mRNA (16 h) expression by RT-PCR analysis as described in the “Methods” section. c Cells were transiently transfected with pGL-MMP9-Luc and pGal for 24 h, pretreated with TSIIA (10 μM), NAC (5 mM), Apo (10 μM), PD98059 (PD, 10 μM), and Rott (1 μM) for 1 h and then stimulation with PMA (1 μM) for 16 h. After stimulation, luciferase activity of MMP-9-promoter construct was measured as relative promoter activity to that of β-galactosidase. Data are expressed as the mean ± SEM (N = 3). ^#P < 0.01, as compared with the respective values of untreated control. *P < 0.05; **P < 0.01, as compared with the respective values of cells stimulated with PMA only. The image represents one of three individual experiments