Figure 4.

Treatment with PEX-9 inhibitor resulting in decreased interaction between MMP-9 and α4β1 integrin and attenuated downstream signaling. (A) Co-IP with HT1080 cells transiently overexpressing MMP-9/MYC cDNA was performed in the presence of 50 μM compound 3c. MMP-9/MYC was immunoprecipitated using an anti-Myc antibody and served as an input control while α4 and β1 integrin interactions were probed via immunoblot. A decrease in MMP-9 interacting with both integrin subunits was observed as well as a (B) decrease in Src^Tyr418 and downstream target proteins FAK^Tyr 576/577/PAX^Tyr118 phosphorylation. (C) 60× confocal images were captured, and representative images of MMP-9 (green) and each integrin subunit (red) were taken after treatment of HT1080 cells with 50 μM of either compound 1a, 3c, or 1f. Scale bar = 100 μm. (D) 60× confocal images were captured, and representative images of p-FAK^Tyr 576/577 (red) and p-PAX^Tyr118 (green) were taken after treatment of HT1080 cells with 50 μM of either compound 1a, 3c, or 1f. Scale bar = 100 μm.