Fig. 5: Combined inhibition of DNA methylation and histone deacetylation decreases proliferative capacity and tumorigenesis of HN9-R clones.

A Colony formation analysis of 5-Aza-treated HN9 cells with or without MS275. B, C In vivo tumorigenesis of HN9-R clones (B) and tumor weight and size of HN9-R xenograft tumors (C) after treatment with 5-Aza and/or MS275. D Immunohistochemistry and the quantitative analysis of Ki67, p62, and H3K27me3 expression in response to treatment with 5-Aza and/or MS275. Scale bars, 50 μm. E, F ChIP analysis of the p62 promoter performed with antibodies against HDACs, DNMT1, p300, and H3 histone modification in HN9-R xenograft tumor tissues treated with 5-Aza and/or MS275. Values are means ± standard deviation (SD) for eight mice (B–F). *p < 0.05; **p < 0.01; ***p < 0.005.