Fig. 4

Knockdown of CD73 in co-culture or fibroblasts reduced of MMP-2 production from fibroblasts. a CD99 siRNA (0.02 pmol/μl) treatment causes knockdown of CD99 expression (32 and 25 kDa) in co-culture of tumor cells (FU-EPS-1) with fibroblasts (ST353i). Protein was extracted from membrane preparation. No reduction of MMP-2 production in conditioned medium in co-cultures of tumor cells with fibroblasts was observed upon siRNA mediated CD99 knockdown in co-cultured cells. b CD73 siRNA (0.01 pmol/μl) treatment causes knockdown of CD73 expression in co-culture of tumor cells (FU-EPS-1) with fibroblasts (ST353i). Protein was extracted from whole cell lysate. Reduction of MMP-2 production in conditioned medium was observed upon siRNA mediated CD73 knockdown in co-cultures of tumor cells with fibroblasts. c Enzyme-linked immunosorbent assay (ELISA) of MMP-2 in conditioned medium of fibroblasts, tumor cells, and co-cultured cells. These co-cultured cells were transfected with CD73-specific or control siRNA. MMP-2 production from co-cultured cells decreased significantly in CD73 transfected cells. *p < 0.01