Figure 5

KDM1 blocker NCL-1 promotes inhibitory epigenetic modifications. (A) Model cells were treated with or without KDM1 inhibitor (NCL-1) and cell proliferation was determined at indicated concentrations using the Cell Titer Glo assay. (B) MCF-7-PELP1 cells were treated with or without NCL-1 (10 μM), and chromatin immunoprecipitation (ChIP) analysis was performed using H3K4me2-specific, H3K9me2-specific or H3K9ac-specific antibodies and the status of epigenetic modifications was analyzed using real-time PCR with aromatase P1.3/II promoter-specific primers. (C) MCF-7-HER2 cells were treated with or without NCL-1 (10 μM), and ChIP analysis was performed using H3K4me2-specific, H3K9me2-specific or H3K9ac-specific antibodies and the status of epigenetic modifications was analyzed using real-time PCR with estrogen receptor target gene GREB1C proximal promoter-specific primers. (D) MCF-7-PELP1 and MCF-7-HER2 cells were treated with NCL-1, total RNA was isolated and expression of aromatase and GREB1 genes was analyzed by quantitative RT-PCR. Error bars indicate ± standard error of the mean. Statistical significance determined by Student's t test. ***P < 0.001, **P < 0.01, *P < 0.05. KDM1, lysine-specific histone demethylase 1; NCL-1, N-((1S)-3-(3-(trans-2-aminocyclopropyl)phenoxy)-1-(benzylcarbamoyl)propyl)benzamide; PELP1, proline glutamic acid and leucine-rich protein 1.