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Figure 2 | Molecular Cancer

Figure 2

From: Epigenetic repression of ROR2 has a Wnt-mediated, pro-tumourigenic role in colon cancer

Figure 2

Promoter methylation-dependent ROR2 repression in colon cancer. (A) qRT-PCR analysis of ROR2 mRNA relative to GAPDH in three samples with unmethylated ROR2 (white) and four samples with ROR2 promoter hypermethylation (red; top panel). The bottom panel shows WB analysis of ROR2 and actin in normal colon epithelium. Two samples show unmethylated ROR2 (HCT116, SW480) and four samples show ROR2 promoter hypermethylation (HT29, HCT15, DLD1, RKO). (B) Relative ROR2 mRNA levels in two cancer cell lines with unmethylated ROR2 (white) and three samples with ROR2 promoter hypermethylation (red) after treatment with the demethylating drug 5-aza-2'-deoxycytidine. (C) qRT-PCR analysis of ROR2 mRNA in 20 primary colorectal tumours and the corresponding normal colon epithelium. Results are shown as the mean ± SD of three independent experiments. (D) Immunohistochemistry analysis of ROR2 in formalin-fixed, paraffin-embedded tissues. The images show protein expression in colorectal gland mucosa (arrow) in normal colon epithelium and lack of ROR2 expression in a poorly differentiated colon adenocarcinoma. (E) Relationship between ROR2 expression and promoter hypermethylation. qRT-PCR quantification of ROR2 relative to GAPDH expression in three ROR2-expressing tumours (U1-3) and three tumours with low ROR2 expression (M1-3; left panel). MSP analysis of ROR2 promoter methylation. A PCR band in lanes M or U indicates methylated or unmethylated, respectively. IVD is used as a positive control for methylated DNA and normal lymphocytes (NL) as negative control (upper right). WB analysis of ROR2 protein expression in the same tumour samples (lower right).

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