Figure 5.

Effect of tissue inhibitor of metalloproteinase‐2 (TIMP‐2) on syndecan‐1 shedding by membrane‐type (MT) 1‐matrix metalloproteinase (MMP). (a) Expression plasmid for syndecan‐1 was cotransfected with control plasmid (lane –) or MT1‐MMP plasmid and indicated amount of TIMP‐2 plasmid into 293T cells, and the culture medium was replaced with serum‐free Dulbecco's Modified Eagle Medium (DMEM) at 48 h after transfection. Medium was harvested after 12 h, and was analyzed for shed syndecan‐1 by dot blot analysis using anti‐syndecan‐1 antibody. (b) Cells cotransfected with syndecan‐1 and MT1‐MMP plasmids were cultured in serum‐free DMEM in the presence or absence of TIMP‐1 or TIMP‐2 (1 µg/mL), and shed syndecan‐1 was analyzed as above. The data represent one of three different experiments showing similar results. The highest shed syndecan‐1 level for each experiment was arbitrarily set to 1, and the levels of other transfections were adjusted accordingly.