Gls2 promoter region was hypermethylated in human liver and colon cancers. The Gls2 promoter methylation status in cancer cells was analyzed by MSP (A) and BGS (B-C). For BGS, each circle indicates one CG site and circles filled in black represent methylated CG sites. One row of circles represents a single colony (B) and a chromatogram representative of BGS results in MHCC-97H and normal liver tissue was also shown (C). USP, unmethylation-specific PCR; Normal 1, normal liver tissue; Normal 2, normal colon tissue. (D) Gls2 promoter methylation status in cancer cells after demethylation treatment. (E) The promoter methylation status of Gls2 in 20 pairs of human HCC tissues and adjacent non-tumor tissues was determined by MSP. (F) Gls2 mRNA level in these tissues was measured by real-time PCR. GAPDH was used to normalize the template amount. USP, unmethylation-specific PCR. “T” indicates tumor tissues and “N” represents adjacent non-tumor tissues.