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Proteins were transferred onto a 0.2 μm pore size nitrocellulose membrane (Novex-Life Technologies, Thermo Scientific) at 10 V for 2 h using 25 mM Tris, 190 mM glycine 20% methanol as transfer buffer

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Metalloproteinase regulation studies document MMP1 and MMP2 gene expression increases at 0.01 nM with TIMP1 increases at all tested concentrations (0.01-100 nM)