Skip to main content

Advertisement

Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Fig. 6 | BMC Cancer

Fig. 6

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. 6

Gö6976 but not Gö6983 strongly inhibits anchorage-independent growth and migration and reverses the E to N-cadherin switch in M4T2 metastatic melanoma cells. a. M4T2 metastatic melanoma cells were seeded in methylcellulose (1000 cells per plate) containing 1 μM Gö6976 or 1 μM Gö6983. After three weeks of culture, the colonies were counted, photographed and their volume determined. The quantifications presented are the means ± SD for three independent experiments. ****, p < 0.0001 versus control DMSO treated cells. b. Scratch wounds were created by scraping confluent monolayers of M4T2 cells, pre-treated one day before with 1 μM Gö6976, 1 μM Gö6983 or DMSO. After 16 and 24 h, cell migration from the wound edges into the wounded area was evaluated. Cells were photographed at 0, 16 and 24 h after wound. The photographs presented are those of a representative experiment. c. Cells were incubated in RPMI medium containing 1 μM Gö6976, 1 μM Gö6983 or DMSO in the upper well of a transwell chamber while in the lower well RPMI medium supplemented with 10% FBS was added. After 24 h, cells that invaded to the lower well were counted. The results presented are the means ± SD for three independent experiments. *, p < 0.05; **, p < 0.01 versus control DMSO treated cells. d. Cells were seeded in 6-well plates at the density of 25,000 cells per well. After three to five days of culture, cells were treated with 1 μM Gö6976, 1 μM Gö6983 or DMSO for 24 h). Cells were then lysed and proteins analyzed by western blot using anti-E-cadherin, anti-N-cadherin or anti-actin antibodies. The autoradiograms presented are those of a typical experiment. The histograms represent quantitative analysis of E- or N-cadherin expression normalized to control DMSO treated cells. The results presented are the means ± SD for two independent experiments. **, p < 0.01; ****, p < 0.0001 versus DMSO treated cells

Back to article page