Fig. 6

Bidirectional regulation between CDH11 and C12orf59. a Schematic illustration of C12orf59 promoter. One potential NF-κB p65 binding sites was shown. b ChIP analysis of NF-κB p65 binding on C12orf59 promoter. ChIP DNA was quantified by real-time PCR primers specific to human C12orf59 promoter. The following PCR primers for ChIP assays were used: 5′- GGAGATCGAGACCATTCTGG-3′, 5′- AAACTCTGCCTCCTGGGTTC-3′. c Fragments encompassing the putative C12orf59 promoter region were inserted upstream of firefly luciferase coding sequences in the pGL3-basic plasmid. The effect of NF-κB protein on transcriptional activity of C12orf59 promoter was measured by luciferase reporter assays. d Overexpression of CDH11 increased C12orf59 mRNA and protein level, while inhibition of NF-κB p65 decreased C12orf59 mRNA and protein level in GC cells. e Molecular mechanism of the C12orf59/ NF-κB/CDH11 feedback loop in promoting GC metastasis