FIGURE 5.

Epstein‐Barrvirus(EBV) factor LMP2A regulated EHF and FZD5 with STAT3 phosphorylation. A, RNA‐seq by EBV factor overexpression in MKN7_WT EHF and FZD5 was highest in LMP2A overexpression compared with the vector control. B, Western blot for p‐STAT3. The protein level of p‐STAT3 was higher in EBV‐positive gastric cancer (GC) than EBV‐negative GC. ATCB was used as a loading control. C, Histone modification states around the EHF promoter and enhancers. Active H3K27ac peaks with the STAT3 motif were observed around EHF in MKN7_WT and MKN7_EB. D, ChIP‐PCR of p‐STAT3 and STAT3 at EHF promoter and enhancers with STAT3 motif. STAT3 and p‐STAT3 are significantly enriched at EHF promoter and enhancer regions in MKN7_EB. E, Knockdown analysis of STAT3. STAT3 knockdown was performed using two different siRNAs (siSTAT3#1 and #2) in MKN7_LMP2A, MKN7_EB, SNU719, YCC10, and NCC24. STAT3, EHF, and FZD5 expression levels were determined by RT‐qPCR, respectively. GAPDH was used as a normalized control for the RT‐qPCR assay. STAT3 knockdown induced EHF and FZD5 downregulation in EBV‐positive GC