The expression of ATP4B and ATP4A and histone modifications in human GC cells upon epigenetic reagent treatment. (A) RT-qPCR analysis of the change of ATP4B expression following treatment with epigenetic reagents, 5-AZA and/or TSA (5-AZA: 5 μM, 96 h; TSA: 5 μM, 24 h). The relative expression of ATP4B mRNA in each cell line was normalized against GAPDH control using the 2−ΔΔCt and further compared to its untreated control. Results shown represent the mean ± SD. 5-AZA, 5-aza-2′-deoxycytidine; TSA, trichostatin A. (B) RT-PCR of ATP4A expression in human GC cell upon epigenetic reagent treatment. (C) Western blotting of BGC823 cells after 5-AZA or TSA treatment. (D) ChIP was performed on BGC823 cells treated with 5-AZA or TSA or DATS (40 μM, 12 h) using antibody against H3K9ac or control IgG. Precipitated ChIP DNA fractions were analyzed by qPCR for the enrichment of H3K9ac in the ATP4B intragenic region. Results are expressed as the percentage of input. DATS, diallyl trisulfide.