Figure 2.

HDACi enhance expression of ERβ target genes. GBM cells (U251 and U87) were treated with vehicle or panobinostat (12.5 nM for U251, 50 nM for U87) or romidepsin (6.25 nM) for 48 h and the status of activation mark H3K9-Ac at the 0N promoter of ERβ (A) and ERβ target genes (B) were analyzed by ChIP. (C) U251-ERβ cells stably expressing ERE-luc reporter and U87-ERβ cells transiently transfected with ERE-luc reporter were treated with LY500307 (10 nm) or panobinostat (6.25 nm for U251, 50 nm for U87) or romidepsin (3.1 nm for U251, 6.25 nM for U87) and the reporter activity was measured after 24 h. U251 (Pano 12.5 nM, Romi 6.25 nM) and U87 (Pano 50 nM, Romi 6.1 nM) (D) and primary GSC-111010 (Pano 100 nM, Romi 25 nM), GSC-012015 (Pano 50 nM, Romi 20 nM), and GSC-101310 (Pano 50 nM, Romi 6.25 nM) (E) cells were treated with panobinostat and romidepsin for 24 h, and the status of ERβ target genes was determined using RT-qPCR. Data are representative of 3 independent experiments (n = 3). Data are represented as mean ± SEM. *P < .05; **P < .01; ***P < .001; ****P < .0001. Statistical differences were examined using Student’s t-test way or 2-way ANOVA. ANOVA, analysis of variance; ChIP, chromatin immunoprecipitation; ERβ, estrogen receptor β; GBM, glioblastoma; HDACi, histone deacetylase inhibitors.