Fig. 4: linc00976 acts as a sponge for miR-3202.

a–c lncLocator, nuclear-cytoplasmic fractionation, and ISH assays revealed the subcellular location of linc00976 in CCA cells. d RIP assay for linc00976 levels in HuCCT1 cells transfected with Ago2-overexpression or control vectors. e Schematic illustration showing overlapping target miRNAs of linc00976 predicted by LNCediting, RegRNA2, and RNA22. f Lysates prepared from CCA cells transfected with linc00976-overexpression vectors or control vectors were subject to RNA pull-down assay, and the efficiency was confirmed by RT-qPCR. g The relative levels of five miRNA candidates in the CCA cell lysates were examined by RT-qPCR. h The putative binding sites between miR-3202 and linc00976. i The luciferase activities was detected in CCA cells after co-transfection with linc00976-WT or linc00976-MUT and miR-3202 mimics or NC mimics. j Anti-Ago2 RIP assay was conducted in CCA cells after transfection with miR-3202 mimics or NC mimics, followed by western blotting and RT-qPCR analyses to detect the expression levels of Ago2, linc00976, and miR-3202. k Relative expression level of miR-3202 in CCA cells and human intrahepatic biliary epithelial cells. n.s. not significant, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. l The expression level of miR-3202 was detected by qRT-PCR in 50 paired CCA tissues and adjacent non-tumor tissues. m Pearson correlation analysis of linc00976 and miR-3202 expression in 50 CCA tissues. CCA cholangiocarcinoma, ISH in situ hybridization, RT-qPCR reverse transcription-quantitative PCR.