PAX2 and ADAM10 expression in melanocytes, keratinocytes and melanoma cells.

<p>(<b>A</b>) Western Blot analysis was performed to determine the PAX2 and ADAM10 expression in melanocytes (Mel43), keratinocytes and melanoma cells (A375, G361, IPC298, MeWo, NW1539 and SKMel5). Notably, 5 of 6 melanoma cell line show PAX2 and ADAM10 expression. β-Actin Western Blot analysis was performed to control equal protein loading. (<b>B</b>) Immunofluorescence staining of primary melanocytes Mel43 (left image) and the melanoma cell lines IPC298 (middle image) and G361 (right image) was performed to investigate the localisation of ADAM10 and PAX2. Cells were incubated with monoclonal ADAM10 and polyclonal PAX2 specific antibodies, followed by Alex488 coupled secondary antibodies (green) and Cy3 coupled secondary antibodies (red). The cells were stained with DAPI to visualize nuclei (blue). (<b>C</b>) The relative immunofluorescence intensity of ADAM10 and PAX2 expression in the melanocytes Mel43 and the melanoma celllines IPC298 and G631 were determined and depicted in a graph. ***P<0.001 PAX2 immunofluorescence intensity considered statistically significant compared to the PAX2 immunofluorescence intensity of melanocytes. <b>###</b>P<0.001 ADAM10 immunofluorescence intensity considered statistically significant compared to ADAM10 immunofluorescence intensity. (<b>D</b>) The specificity of ADAM10 and PAX2 immunofluorescence staining was controlled by using isotype specific control (control IgG) antibodies.