Fig. 4

HNF4A transcriptionally regulates MIR194-2HG expression. (A, B) The knockdown and overexpression efficiency of HNF4A were determined in GC cell lines by qRT-PCR assay. (C) Western blotting assays confirmed that the gain-of function and loss-of-function studies of HNF4A were successful. (D, E) Quantitative RT-PCR assay showed that MIR194-2HG was downregulated in HNF4A-silenced GC cells but upregulated in HNF4A overexpression GC cells. (F) Promoter analysis showed that MIR194-2HG contained an obvious HNF4A binding motif. (G) Chip-seq analysis showed that HNF4A directly binds to MIR194-2hg promoter in liver and colon cells. (H) Chip-qPCR assays showed that HNF4A directly binds to the promoter of MIR194-2HG. The Ubiquitin gene promoter DNA was used as an internal control. (I) MIR194-2HG was highly co-expressed with HNF4A in colon, stomach, and small intestine tissues