Figure 3. MeCP2 regulates GPC5-AS1 expression by binding its promoter regions. (A) MeCP2 binding sites were relative to the CpG island location. (B, C) ChIP RT-PCR of GPC5-AS1 was performed with an anti-MeCP2 antibody. (D) ChIP RT-PCR of GPC5-AS1 was performed with an anti-GFP antibody after transfection with the GFP-MeCP2 plasmid. (E) ChIP RT-PCR of GPC5-AS1 was performed with an anti-GFP antibody after transfection with Ctrl (GFP plasmid), WT (GFP-MeCP2 plasmid), MT1 (GFP-Mutation type 1), and MT2 (Mutation type 2). (F) SGC-7901 cells were transfected with pGL3-GPC5-AS1-luc and pGL3-GPC5-AS1-luc + Methylation; luciferase activity was determined at 48 h post-transfection. Renilla luciferase served as the internal control. (G) SGC-7901 cells were treated with pGL3- GPC5-AS1-luc, pGL3-GPC5-AS1-luc + Methylation, MeCP2 siRNAs, and overexpression vectors; luciferase activity was determined. (H) Correlation analysis of MeCP2 and GPC5-AS1 expression in GC tissues through starBase (r = -0.192, p < 0.05). (I) Expression of GPC5-AS1 in SGC-7901 and BGC-823 cells transfected with si-MeCP2. (p* < 0.05, p** < 0.01, p*** < 0.001).