VGF expression vector (pCMV6-AC–VGF–GFP, OriGene Technologies, Inc., Rockville, MD), as well as the empty vector (pCMV6-AC-GFP, OriGene Technologies, Inc.)were transfected into J82 and SCaBER bladder cancer cell lines that have fully methylated VGF promoter of our analyzed region and VGF silenced. Here, VGF is linked to urinary bladder cancer.