Fig 1: LncRNA KCNQ1OT1 increased EIF2B5 promoter methylation by recruiting DNA methyltransferases into its promoter region. A, B RT-qPCR analysis and Western blotting for the mRNA and protein levels of DNMT1, DNMT3A and DNMT3B in SKOV3 and SW626 cells transfected with Control (blank), sh-NC or sh-KCNQ1OT1. C Prediction for the binding potency between KCNQ1OT1 and DNMT1, DNMT3A or DNMT3B. D RIP assay for the combining ability of KCNQ1OT1 to DNMT1, DNMT3A and DNMT3B. E ChIP assay for the enrichment of DNMT1, DNMT3A and DNMT3B in EIF2B5 promoter. *P < 0.05
Fig 2: Increased expression of lncRNA KCNQ1OT1 and decreased expression of EIF2B5 in OC. A RT-qPCR analysis for the enrichment of KCNQ1OT1 and EIF2B5 in OC tissues and normal tissues (N = 32). B Spearman’s correlation analysis for the correlation between expression of KCNQ1OT1 and EIF2B5 in OC tissues (N = 32). C RT-qPCR analysis for the enrichment of KCNQ1OT1 and EIF2B5 in IOSE-80, A2780, Anglne, SKOV3 and SW626 cells. D Western blotting for abundance of EIF2B5 protein in IOSE-80, A2780, Anglne, SKOV3 and SW626 cells. ***P < 0.001. **P < 0.01. *P < 0.05
Fig 3: EIF2B5 inhibition largely relieved the inhibitory impact of KCNQ1OT1 depletion on the malignant behaviors of OC cells. SKOV3 and SW626 cells were transfected with Control (blank), sh-KCNQ1OT1, sh-EIF2B5 or sh-KCNQ1OT1 + sh-EIF2B5. A, B RT-qPCR analysis and Western blotting for the mRNA and protein levels of EIF2B5 in transfected cells. C MTT assay for the cell viability of transfected cells. D Colony formation assay for the cell clonogenicity capacity of transfected cells. E Wound healing assay for the cell migration of transfected cells. F Transwell assay for the cell invasion of transfected cells. G Western blotting for abundance of E-cadherin and N-cadherin proteins in transfected cells. *P < 0.05
Fig 4: LncRNA KCNQ1OT1 could decrease EIF2B5 expression by promoting EIF2B5 methylation. A, B RT-qPCR analysis and Western blotting for the mRNA and protein levels of EIF2B5 in SKOV3 and SW626 cells transfected with Control (blank), sh-NC or sh-KCNQ1OT1. C Nuclear-cytoplasmic fractionation assay for the subcellular location of KCNQ1OT1 in SKOV3 and SW626 cells, GAPDH and U6 snRNA serving as internal control for cytoplasmic and nuclear cytoplasmic and nuclear fractions, respectively. D FISH analysis for the subcellular location of KCNQ1OT1 in SKOV3 and SW626 cells. E Prediction for the CpG island in EIF2B5 promoter region by MethPrimer 2.0. F MS-PCR assay for the methylation status of EIF2B5 promoter in SKOV3 and SW626 cells transfected with Control (blank), sh-NC or sh-KCNQ1OT1; M: Methylated primers, U: unmethylated primers. *P < 0.05
Supplier Page from Abcam for Anti-EIF2B5 antibody [EPR13533]