Fig. 3
Epigenetic inhibitors fail to fully restore gluconeogenic gene expression with or without glucagon. a Diagram of epigenetic drugs and their targets. GSK126: EZH2 (enhancer of zeste homolog 2) inhibitor, SAM: S-adenosyl-L-methionine, SAH: S-adenosyl homocysteine, H3K27me3: histone 3 lysine 27 trimethylated, LBH589: pan-HDAC (histone deacetylase) inhibitor, H3K27Ac: histone 3 lysine 27 acetylated, Decitabine: DNA methylation (DNMT -DNA methyltransferase) inhibitor, 5mC: 5-methylcytosine. b Protein analysis of epigenetic inhibitor efficacy in SNU398 eGFP-expressing cells at the indicated drug concentrations, culture conditions, and time. c qPCR mRNA levels of gluconeogenic genes in SNU398 expressing eGFP and treated with epigenetic inhibitors at the concentrations used in b. Data represent a single experiment with 2 biological replicates (2 separate RNA samples). ND: not detected. d qPCR mRNA levels of G6PC, FBP1, and PCK1 in SNU398 GCGR-overexpressing cells treated with glucagon plus combinations of epigenetic drugs. Data represent a single experiment with 3 technical replicates (1 RNA sample). Error bars: ±SD. (−): no drug, 100 G: 100 nM glucagon, vehicle: 0.035% of 0.05 M acetic acid, EZH2i: 1 μM GSK126, HDACi: 10 nM LBH589, DNMTi: 5 μM Decitabine, triple: 1 uM GSK126 + 10 nM LBH589 + 5 uM Decitabine. e Same qPCR mRNA analysis as in d but using the liver cancer patient-derived cell line, M7571