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. 2012 Nov 23;10:35. doi: 10.1186/1478-811X-10-35

Figure 3.

Figure 3

BK induces MMP-9 expression via p47phox translocation in RBA-1 cells. (A) Cells were pretreated without or with Apocynin (Apo, 10 μM) for 1 h before exposure to 10 nM BK for the indicated time intervals. The conditioned media were collected for zymographic analysis of MMP-9 expression. (B) Cells were pretreated with or without Apo (10 μM) for 1 h before exposure to 10 nM BK for 16 h. The total RNA was collected and analyzed by RT-PCR. (C) Cells were pretreated without or with Apo (10 μM) for 1 h treated with 10 nM BK for the indicated time intervals or 3 min. The membrane and cytosol fractions were prepared and analyzed by Western blotting. The p47phox translocation was also confirmed by immunofluorescent staining. (D) Cells were pretreated without or with Apo (10 μM) for 1 h before exposure to 10 nM BK for 5 min. The Nox activity and ROS generation were analyzed. (E) Cells were transfected with scramble (scra) or p47phox siRNA for 24 h, followed by incubation with 10 nM BK for 24 h. The conditioned media and cell lysates were collected for zymography of MMP-9 or Western blotting to determine the levels of Nox2 and GAPDH (as an internal control). Data are expressed as the mean ± SEM (n = 3). *P < 0.05; #P < 0.01, as compared with the respective values of cells stimulated with vehicle (C) and BK (A, B, D, E) only. The figure represents one of three similar experiments.

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