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. 2017 Nov 6;37(6):BSR20171070. doi: 10.1042/BSR20171070

Figure 4. DANCR epigenetically suppresses lncRNA-LET expression through association with EZH2 and HDAC3.

Figure 4

(A) The DANCR-overexpressed AGS cells were treated with 5 μM DZNep and/or 1 μM SAHA for 48 h, and the relative expression of lncRNA-LET was detected by qPCR. (B) The DANCR-overexpressed AGS cells were transfected with EZH2 and/or HDAC3. After 48 h, the relative expression of lncRNA-LET was detected by qPCR. (C) DANCR RNA levels in immunoprecipitates by EZH2 or HDAC3 were determined by qPCR. DANCR RNA expression levels are presented as fold enrichment values relative to IgG immunoprecipitates. (D) EZH2 and HDAC3 protein levels in immunoprecipitates with biotin-labeled DANCR RNA were evaluated by Western blot. (E) The occupancy level of EZH2, HDAC3, H3K27me3, H3Ac, and H4Ac at lncRNA-LET promoter region was determined by ChIP assay and followed by qPCR in control and DANCR-silenced BCG-823 cells. (F) The occupancy level of EZH2, HDAC3, H3K27me3, H3Ac, and H4Ac at lncRNA-LET promoter region was determined by ChIP assay and followed by qPCR in control and DANCR-overexpressed AGS cells. (G) BCG-823 cells were transfected with DANCR siRNAs for 48 h. After immunoprecipitating endogenous EZH2, bound HDAC3 was subjected to Western blotting. All experiments were repeated three times. Data are shown as mean ± SD; *P<0.05.