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Fig. 2 | Molecular Cancer

Fig. 2

From: Wnt-signaling enhances neural crest migration of melanoma cells and induces an invasive phenotype

Fig. 2

Wnt3a enhances and PKF115–584 blocks neural crest migration of SKMEL28 melanoma cells in the chick embryo in vivo. a Control treated (n = 8), Wnt3a (n = 10) or 0.5 μM PKF115–584 (n = 12) pre-conditioned SKMEL28 melanoma cell aggregates were injected into the neural tube of stage 12/13 HH chick embryos and further incubated for 48 h. Embryos were formalin-fixed and paraffine-embedded (FFPE) before sectioning and staining for HMB45 (red) to identify the SKMEL28 melanoma cells. In 6/8 embryos, untreated HMB45-positive SKMEL28 cells integrated into the roof plate of the neural tube and migrated mainly along the proximal medial neural crest cell pathway. Wnt3a pre-conditioned SKMEL28 cells similarly integrated into the roof plate and predominantly performed medial neural crest migration in 8/10 embryos (not significant, Fisher’s Exact Test, compared to the untreated cells). An increased number of HMB45-positive Wnt3a pre-conditioned cells were detected along the distal medial neural crest cell pathway close to the notochord and the developing para-aortic sympathetic ganglia. 0.5 μM PFK115–584 pre-conditioned, HMB45-positive SKMEL28 cells formed compact tumor nodules in the dorsal lumen of the neural tube and were attached to and integrated into the roof plate and the adjacent mesenchyme between the roof plate and the surface ectoderm. Only in 1/12 embryos single emigrating cells were detected along the proximal medial neural crest cell pathway in direct vicinity to the lateral roof plate (p < 0.01, Fisher’s Exact Test, compared to the untreated cells). b Scheme showing the neural crest-like migration patterns of melanoma cells out of the roof plate after injection into the neural tube. c Quantification of chick embryos showing SKMEl28 melanoma cells that migrated to proximal or distal sites of the medial migration pathway after different pre-treatments (asterisks define p-values calculated by Fisher’s exact test)

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