Reactivation of CTBP2 expression by EZH2i and HDACi in MM. (A) Integration of ChIP-seq data from the BLUEPRINT Consortium and the ENCODE project in the UCSC genome browser for H3K27me3 levels around the CTBP2 locus at the stages of NPC development and MM. (B-C) Restoration of CTBP2 expression in HMCLs. (B) RQ-PCR of HMCLs treated with a serial dose of DZNep or TSA. GAPDH served as an internal control. Ordinary 1-way ANOVA followed by Dunnett multiple comparison test. (C) Immunoblotting of HMCLs treated with (left) DZNep at 10 μM and (right) TSA at 500 nM. β-actin served as the loading control. (D-F) HMCLs were treated with a single agent or combination of EZH2i and/or HDACi. Expression of CTBP2 (upper) and MYC (middle) as evaluated by RQ-PCR and cell proliferation (lower) using the CellTiter-Glo assay. Ordinary 1-way ANOVA followed by Tukey multiple comparison test. (D-E) MM.1S, NCI-H929, OPM2, and RPMI8226 were treated with DZNep and/or TSA at indicated dose. Expression changes in CTBP2 and MYC are illustrated by (D) RQ-PCR and (E) immunoblotting. (F) Treatment of EPZ-6438 (EPZ) at the indicated dose and/or panobinostat (Pano) at 10 nM in MM.1S and NCI-H929. Expression changes in CTBP2 and MYC are illustrated by (upper) RQ-PCR and (lower) immunoblotting. The results are expressed as the mean ± SD of triplicate measurements from at least 3 independent experiments. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001, ∗∗∗∗P < .0001; ns, not significant.