1. Effect of G9a on the expression of GSH synthesis enzymes.
Effect of G9a on the expression of GSH synthesis enzymes. A, Quantitative real-time PCR analysis of indicated mRNA expression in SAS and SAS-CR cells (top), SAS-CR cells expressing control or shG9a plasmid (middle), and SAS-CR cells treated with vehicle or 1 μmol/L UNC0638 for 24 hours (bottom). All values are the means of three independent experiments. Bar: mean ± SD. B, Western blot analysis of indicated protein expression in SAS, SAS-CR cells and SAS-CR cells expressing shLuc or shG9a, or treated with DMSO or UNC0638 (1 μmol/L) for 24 hours. A blot representative of results obtained in three independent experiments is shown. C,GCLC promoter reporter activity of SAS and SAS-CR cells (top), or SAS-CR cells treated with vehicle or 1 μmol/L UNC0638 for 24 hours (bottom). D, ChIP assay analysis of the binding of indicated proteins to the GCLC promoter in SAS-CR cells. All values are the means of at least three independent experiments. Bar: mean ± SD. E, ChIP assay analysis of binding of ATF4 to the AP1-2 site of GCLC promoter and binding of Nrf2 to the ARE3 or ARE4 site in SAS-CR cells treated with vehicle or 1 μmol/L UNC0638 for 24 hours. All values are the means of at least three independent experiments. Bar: mean ± SD. F, Lysates of SAS-CR cells were prepared for immunoprecipitation with anti-H3K9me1 (left) or anti-ATF-4 (right) antibodies followed by Western blot analysis of indicated proteins. A blot representative of results obtained in three independent experiments is shown in each case (*, P < 0.05; **, P < 0.01; ns, not statistically significant).
Chia-Wen Liu et al. Mol Cancer Ther 2017;16:
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