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. 2012 Sep 18;287(45):37732–37744. doi: 10.1074/jbc.M112.389015

FIGURE 6.

FIGURE 6.

MMP and ERK activity are required for eHsp90-mediated motility and EMT events. A, a gelatin zymography assay was utilized to assay MMP-2/9 activity in control or ARCaPE-Hsp90 cells. For the indicated inhibitors, cells were treated for 2 days prior to media collection. Cells were treated with ERK inhibitor (UO126, 10 μm), MMP-2/9 inhibitor (SB-3CT, 1 μm), or NPGA (1 μm). B, transcript expression of E-cadherin was evaluated in ARCaPE-eHsp90 following a 3-day treatment with the following: NPGA (1 μm), pan-MMP inhibitor (GM6001, 1 μm), MMP-2/9 inhibitor (SB-3CT, 1 μm), MMP-3 inhibitor (inhibitor IV, 5 μm), or ERK inhibitor (UO126, 10 μm). C, immunoblot analysis of E-cadherin and ERK proteins in ARCaPE-eHsp90 following the time-dependent inhibition of the following: MMP-2/9, MMP-3, or ERK. D, the effect of 3 days of MMP and ERK inhibition upon E-cadherin and ZO-1 localization in ARCaPE-LacZ and ARCaPE-eHsp90 was assessed by confocal microscopy. Cells were treated as in A, with inclusion of the pan-MMP inhibitor (GM6001, 1 μm) and the MMP-3 inhibitor (inhibitor IV, 5 μm). E, evaluation of MMP and ERK in directing eHsp90 cell motility following a scratch wound assay. Scale bar is 50 μm. UT refers to untreated vehicle control. Asterisks (*) indicate significance p value ≤0.05.