Figure 4.

LPA induces MMP-9-dependent E-cadherin ectodomain shedding. (a) OVCA429 and OVCA433 cells were treated with LPA (0, 20, or 80 μM, as indicated) and conditioned media evaluated for MMP expression by gelatin zymography. (b) Cells were treated with LPA (20 μM) in the presence or absence of anti-MMP-9 function blocking antibody (10 μg/mL). Conditioned media were subjected to immunoprecipitation with anti-E-cadherin antibodies, and precipitates were western blotted with a second E-cadherin antibody (Zymed, 1 : 1,000) followed by peroxidase-conjugated secondary antibody (1 : 5,000) and enhanced chemiluminescence detection. The lower panel shows densitometric quantitation of replicate western blots.