Fig. 2

Magnolin suppresses EGF-induced cell migration in JB6 Cl41 cells. a Magnolin inhibits COX-2 protein levels. JB6 Cl41 cells were starved, treated with the indicated doses of magnolin and EGF for 30 min, and then the COX-2 protein level was visualized by Western blotting. β-actin was used as an internal control to verify equal protein loading. b–c Magnolin suppressed EGF-induced Cox-2 promoter activity. JB6 Cl41 cells stably expressing Cox-2-promoter luciferase reporter plasmid were seeded, cultured, starved and the luciferase activities analyzed for the Cox-2 promoter activity by combination treatment with EGF and magnolin as indicated (b), or the Cox-2 gene expression was analyzed with magnolin treatment under normal cell culture conditions for 24 h (c) as described in “Materials and Methods”. d Magnolin inhibits EGF-induced cell migration. JB6 Cl41 cells were seeded into culture-inserts and cultured overnight. Cell proliferation was stopped by mitomycin-C treatment for 2 h, and culture inserts were removed to offer a cell-free gap. The cells were treated with EGF or EGF and magnolin for 24 h, cell migration was observed under a microscope, and then the migrated area was measured using the Image J computer software program (v. 1.45). a–d Data are presented as the mean ± S.D. of values from triplicate experiments, and statistical significance was determined using the Student’s t-test (*, p < 0.05)