Figure 4.

Analysis of N-cadherin downstream signaling pathways in MUC4-expressing and control cells. (a) Western blot analysis of N-cadherin downstream signaling molecules showed increased activation of serine (473) Akt, ERK1/2 and increased expression of MMP9 in MUC4-overexpressed cells. No variation was seen in threonine (Y308) phosphorylated Akt. The total form of Akt and ERK1/2 were unchanged in both the SKOV3-derived sublines. β-actin was used as a loading control. (b) Western blot analysis showed that pFAK inhibition also reduces the phosphorylation of serine (473) Akt, ERK1/2 and decreases the expression of MMP9 compared with the mock control. (c, d) Wound healing assay: The scratch was made across the cell monolayer in pFAK (1,2,4,5-benzenetetraamine tetrahydrochloride) and pJNK (SP600125) inhibitor-treated SKOV3-MUC4 cells with mock control. The migration of cells was measured (μm²) in treated and untreated cells. Significant migration of cells was analyzed using two-tailed Student's t-test. A P-value of <0.05 was considered statistically significant. (FAK Inhi-10 mm, *P <0.0004; FAK Inhi-50 μm, *P <0.0003), (JNK1/2 Inhi-20 nm, *P <0.005; JNK1/2 Inhi-40 nm, *P <0.0002; JNK1/2 Inhi-80 nm, *P <0.0001) (original magnification × 100, scale bar-0.8 mm).