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. Author manuscript; available in PMC: 2016 Jul 15.
Published in final edited form as: Cell Rep. 2016 Jun 23;16(2):457–471. doi: 10.1016/j.celrep.2016.05.087

Figure 1. Oncogenic EGFR is necessary for epigenetic silencing of TSGs in lung cancer cells (See also Figure S1).

Figure 1

(A) (Left) Venn diagram shows the number of genes that are commonly or specifically upregulated in HCC827/Del (DEL) or HCC827/Del-TM (DEL-TM) cells after decitabine and vorinostat treatment. (Right) Heatmap of indicated genes in HCC-827/Del (DEL) and HCC827/Del-TM (DEL-TM) cells after decitabine and vorinostat treatment. (B–E) HCC827/Del cells were treated with gefitinib (0.1 μM), afatinib (0.1 μM), or DMSO for 48 h. (B) mRNA expression for the indicated TSGs was measured by RT-qPCR. Gene expression was normalized with respect to GAPDH mRNA expression. Relative expression for the indicated genes in drug-treated cells compared to DMSO-treated cells is plotted. (C) DNA methylation of CpG islands in the promoters of the indicated genes was measured using Me-DIP analysis. The percent of CpG island DNA methylation for the promoters of the indicated genes in drug-treated cells is compared with that of DMSO-treated cells. (D) RNA polymerase II enrichment on TSG promoters was measured via ChIP assay. Relative enrichment in drug-treated cells is compared with that of DMSO-treated cells. (E) Histone H3 lysine 9 acetylation (H3K9-Ac) marks on the indicated gene promoters were measured. Relative amounts of H3K9-Ac marks in drug-treated cells are shown relative to those observed in DMSO-treated cells. (F) The indicated EGFR-mutant lung cancer cell lines were treated with afatinib (0.1 μM) or DMSO for 48 h. mRNA expression levels for the indicated TSGs were measured via RT-qPCR. Gene expression was normalized with respect to GAPDH mRNA expression. Relative expression levels for the indicated genes in drug-treated cells are compared with those of DMSO-treated cells. *p < 0.05.