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Analysis of the smokeless tobacco extracts – induced secretory phenotype. Concentrated conditioned medium from control (Ctr) or tobacco-treated 82.6 and WI-38 fibroblasts were normalized to the cell number and used for gelatin and casein substrate zymography for detection of MMP-2 and MMP-3 activity, respectively, and Western blottingfor MMP-1, MMP-2, and MMP-3 (A). Conditioned medium was also analyzed for IGFBP4, TIMP-2, and TIMP-1 by Western blotting(B).
