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For replicate experiments with numerical values, an analysis of variance was performed using Tukey comparison test. Statistically significant results were indicated (***P? http://www.selleckchem.com/products/PF-2341066.html different controls performed with or without the first antibody on the co-cultured keratinocytes, as well as na?ve keratinocytes, ruled out the non-specific antibody reactions as the source of the signal. The signal-to-noise ratio was 7.10 for co-cultured melanoma cells, 2.04 for co-cultured keratinocytes and 1.05 for na?ve keratinocytes (not shown). Immunolabelling of co-cultured cells http://en.wikipedia.org/wiki/DYRK1B on membranes by melanocyte-, keratinocyte- and melanosome-specific markers confirmed the transfer of melanosomes to keratinocytes and excluded cell passage through membrane pores (Fig.?2). Segregated populations of keratinocytes were observed in one side of the membrane (Fig.?2a�Cf), whereas pure melanocytes were observed on the other side (Fig.?2g�Cl). http://www.selleckchem.com/products/INCB18424.html Interestingly, points of contact between melanoma cells and keratinocytes were visualised at the level of the interconnecting pores of the membranes (Fig.?2m�Cr), suggesting that both cell types were allowed to get in close contact across the membranes. More importantly, transfer of melanosomes, as assessed by HMB-45 immunostaining, was also observed on the keratinocytes layer (Fig.?2c,f). Co-cultures were not damaged during trypsinisation despite the dense incorporation of their processes into membrane pores. This was shown by trypan blue examination of the trypsinised cells showing always