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. 2012 Sep 27;287(46):38957–38969. doi: 10.1074/jbc.M112.357863

FIGURE 5.

FIGURE 5.

Endogenous βig-h3 cleavage by MMP-9 in glioma cells. A and B, βig-h3 and MMP-9 in MMP-9-transfected U87MG cells. A, βig-h3 and MMP-9 expression in the vehicle (vehicle/U87MG)-transfected or MMP-9 (MMP-9/U87MG)-transfected U87MG cells were examined by immunoblotting. B, MMP-9 activity was detected by gelatin zymography. M is a marker for active MMP-9 and pro-MMP-2 (n = 3). C, co-immunoprecipitation of MMP-9 with βig-h3 from the vehicle (vehicle/U87MG)-transfected or MMP-9 (MMP-9/U87MG)-transfected U87MG cells. After the lysates from the vehicle (vehicle/U87MG)-transfected or MMP-9 (MMP-9/U87MG)-transfected U87MG cells were immunoprecipitated (IP) with anti-βig-h3 (+) or control rabbit IgG (−), they were subjected to immunoblotting (IB) using mouse antibody for Myc (MMP-9) (n = 4). D, MMP-9 overexpressed in cells alters cell invasion. Cell invasion in vehicle-transfected, MMP-9-transfected, or IL-1β (10 ng/ml)-treated U87MG cells was examined on Matrigels. βig-h3 and MMP-9 proteins from IL-1β-treated human glioma U87MG cells were examined by immunoblotting and zymography. The control represents untreated samples (n = 3). E, βig-h3 knockdown alters cell invasion. Cell invasion in control siRNA or βig-h3 siRNA-transfected U87MG cells was examined on Matrigels. The βig-h3 and MMP-9 from these cells were examined by immunoblotting and zymography. Control represents control siRNA. (*, p < 0.05; **, p < 0.01, n = 3).