Figure 6

Effect of JAK2 inhibitor on STAT3 activation, EGF-induced N-cadherin, vimentin and cell migration. (A) Expression of P-STAT3 in OVCA 433 cells post-treatment with EGF and EGF+AG490 a specific JAK2 inhibitor. Cells were incubated in normal growth medium (control), medium treated with EGF (10 ng ml⁻¹) or treated with EGF (10 ng ml⁻¹) and AG490 (100 μ M) for 1 or 4 h. respectively. Western blot analysis was performed as described above for P-STAT3 followed by re-probing with STAT3 and β-actin. ^*P<0.05, compared to control untreated cells and, ^§compared to cells in the presence of EGF. Densitometry was performed as described above. (B and C) Expression of N-cadherin and vimentin 24 h post EGF in the absence and presence of AG490 in OVCA 433 cells. Western blot analysis for N-cadherin and vimentin was performed as described above. ^*P<0.05, compared to control untreated cells, and ^§compared to cells in the presence of EGF. (D) OVCA 433 cells were cultured on glass coverslips in either normal growth medium (control) or EGF-treated medium (10 ng ml⁻¹) or in the presence of EGF (10 ng ml⁻¹) and AG490 (100 μ M). Wounding assay was performed as described above. Images are representative of one independent experiment performed three times. Graphs represent mean % of wound closure from three independent experiments. Scale=300 μm. ^*P<0.05, compared to control untreated cells, and ^§compared to cells in the presence of EGF. (E) IL-6 production in the presence and absence of EGF and EGF+AG490 was measured as described above. ^*P<0.05, compared to control untreated cells, and ^§compared to cells in the presence of EGF and AG490.