Figure 4.

The Knockdown of LINC00470 Reverses the Expression of ELFN2 in GBM Cells through an Epigenetic Regulatory Mechanism

(A) The methylation level of ELFN2 was detected by BSP. (B) Western blotting was performed to detect the expression of EED and EZH2 in GBM cells after LINC00470 knockdown. (C) qRT-PCR was performed to detect the expression of ELFN2 in GBM cells after EZH2 knockdown. The data are presented as the means ± SEM of three independent experiments. *p < 0.05. (D) qRT-PCR was performed to detect the expression of ELFN2 in GBM cells after EED knockdown. The data are presented as the means ± SEM of three independent experiments. *p < 0.05. (E) The promoter activity of ELFN2 core promoter constructs was analyzed by luciferase reporter assays. pGL3-control was used as the positive control, and pGL3-enhancer was used as the negative control. The core promoter region is considered to show higher relative luciferase activity than the pGL3-enhancer. The data are presented as the means ± SEM of three independent experiments. **p < 0.01. (F, G, and H) The histone occupancy of the ELFN2 promoter was affected by si-EZH2 (F), si-LINC00470 (G), and si-EED (H). A ChIP assay was performed to detect the H3K4me2, H3K27me3, H3K9me3, and H4K20me3 occupancy of the ELFN2 core promoter. The data are presented as the means ± SEM of three independent experiments. *p < 0.05.