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Fig. 4 | BMC Cancer

Fig. 4

From: Protein kinase C inhibitor Gö6976 but not Gö6983 induces the reversion of E- to N-cadherin switch and metastatic phenotype in melanoma: identification of the role of protein kinase D1

Fig. 4

Gö6976 but not Gö6983 induces rapid β-catenin translocation outside of the nucleus and reduces cyclin D1 expression in M2 metastatic melanoma cells. M2 metastatic melanoma cells were seeded in 6-well plates at the density of 12,500 cells per well. After three to five days of culture, cells were treated with 1 μM Gö6976 or 1 μM Gö6983 for different periods of time (0, 1, 3, 8, 24 or 48 h). a. After treatment, cells were fixed, permeabilized, stained for β-catenin (green) and analyzed under a confocal microscope. Isotype-matched control antibody and DAPI (blue) were used. Images are representative fields from three or more independent experiments. b. After treatment cells were lysed and proteins analyzed by western blot using anti-β-catenin or anti-actin antibodies. The autoradiograms presented are those of a typical experiment. c. After treatment cells were lysed and proteins analyzed by western blot using anti-cyclin D1 or anti-actin antibodies. The autoradiograms presented are those of a typical experiment. The histogram represent quantitative analysis of cyclin D1 expression following Gö6976 treatment normalized to untreated control cells (0 h). The results presented are the means ± SD for three independent experiments. **, p < 0.01; ****, p < 0.0001 versus untreated cells

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