Fig. 1

Evaluation of Sema5A expression in melanoma. a Box and whisker plots showing data from the microarray datasets GDS3964 and GDS1375 downloaded from the Gene Expression Omnibus (GEO, https://www.ncbi.nlm.nih.gov/geoprofiles) website (see Methods for details). Values of Sema5A gene expression levels are reported as counts. Statistical analysis was performed applying Mann-Whitney test. b Western blotting analysis of Sema5A protein expression in melanoma parental cell lines. Reported image is representative of two independent experiments with similar results. β-actin expression was evaluated as control of equivalent transfer and loading. c-g IHC analysis of Sema5A expression in human melanoma specimens. c-e Three representative samples showing different levels of Sema5A selected from metastatic specimens obtained from melanoma patients: c SCORE 1+ (staining that is faint/barely detectable), d SCORE 2+ (staining that is weak/moderate), e SCORE 3+ (staining that is intense/strong). Scale bar 20 μm. f, g Representative images from in situ melanoma specimens showing no immunoreactivity f or focal positivity g for Sema5A expression. Arrow indicates positive cell. Scale bar 30 μm