Figure 3. RHCG is downregulated in HNSCC tissues and cell lines. (A) Western blotting analysis of RHCG protein levels in HNSCC and normal tissues (n=3). (B) mRNA expression of RHCG in HNSCC and corresponding normal tissues (n=20). (C) Expression of RHCG as examined by immunohistochemistry (×400) in in HNSCC and normal tissues. (D) Western blotting analysis of RHCG expression in a human immortalized oral epithelial cell (HIOEC) line and HNSCC cell lines. (E) Real-time PCR analysis of relative RHCG mRNA expression levels HIOEC cells and HNSCC cell lines. (F) Comparison of RHCG expression in cancerous tissues and normal tissues in 4 different cohorts, including Ginos Head-Neck, Peng Head-Neck, Toruner Head-Neck and Ye Head-Neck according to the Oncomine database. (G) Meta-analysis of RHCG expression across the 4 analyses. (H) RHCG protein expressed in normal and HNSCC cancerous tissues from The Human Protein Atlas database. The tissue expressing abundant RHCG signals was normal tongue tissue from a female at an age of 73. RHCG was strongly stained in squamous epithelial cells, mainly located in cytoplasmic or membranous region; the tissue with no staining was head and neck squamous cell carcinoma tissue from an 81-year-old woman.