To verify the robustness of the observed differential methylation, pyroseqeuencing was performed to quantify methylation at the same CpG site of the top differentially methylated gene (HOXA5) and a gene from the top differentially methylated pathway (WNT5A, hedgehog signalling) in an independent series of tumor and matched tumor-adjacent gastric tissues from 60 subjects with GC. The gene discussed is WNT5A; the disease is gastric cancer.