Fig. 4

PCGEM1 directly interacts with miR-182. a Cytoplasmic and nuclear RNA fraction assay showed the cellular location of PCGEM1 in CC cells. U6 RNA served as a positive control for nuclear gene expression. GAPDH mRNA served as a positive control for cytoplasmic gene expression. b MS2-RIP followed by qRT-PCR was conducted to detect endogenous microRNAs associated with lncRNA PCGEM1. c HeLa and SiHa cell lysates were incubated with biotin-labeled wild-type or mutant PCGEM1; after pull-down, microRNAs were extracted, and the amount of miR-182 was assessed by qRT-PCR. d HeLa and SiHa cells were cotransfected with miR-182 mimics and luciferase reporters containing wild-type or mutant PCGEM1. After 48 h, luciferase activity was measured and presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. e Anti-AGO2 RIP was performed in HeLa and SiHa cells transfected with miR-NC or miR-182, followed by qRT-PCR to detect PCGEM1 associated with AGO2. f SiHa cells were transfected with wild-type or mutant PCGEM1. After 48 h, the miR-182 expression was assessed by qRT-PCR. g HeLa cells were transfected with PCGEM1 siRNAs. After 48 h, the miR-182 expression was assessed by qRT-PCR. h The expression of miR-182 in 68 pairs of CC and normal cervical tissues was determined by qRT-PCR. i Expression levels of PCGEM1 and miR-182 were subjected to Pearson correlation analysis. *p < 0.05, **p < 0.01, ***p < 0.001