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Sema4D is highly expressed in HNSCC. (A) RNAs from controls and HNSCC cell lines were compared to reference RNA by microarray analysis. Sema4D mRNA levels are expressed relative to normal human oral keratinocytes (NOK). RNA from brain tissues was used as the positive control. IOK, immortal human oral keratinocytes. (B) Immunoblot analysis for Sema4D shows a band at ≈125 kDa for endogenously expressed protein in lysates from Jurkat cells (positive control) and HNSCC cell lines, but not in the nontumorigenic epithelial-derived cell lines HaCaT or HeLa. Tubulin was used as a loading control. (C) Immunohistochemical analysis of Sema4D in HNSCC. No Sema4D was detected in normal parakeratinized stratified squamous epithelium (Normal) and in dysplastic stratified squamous epithelium, although chronic inflammatory cells were positive (Dysplastic). Sema4D immunoreactivity is seen in invading islands of transformed epithelial cells (Carcinoma). A cell surface-staining pattern is seen at a higher magnification (Insets).

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