Fig. 2

MiR-300 targets to YY1 and inactivates WNT pathway in HCC. a The upstream miRNAs of YY1 were predicted by five programs in starBase (including miRmap, TargetScan, PicTar, miRanda and microT), and the prediction results were showed by Venn. b RIP assay plus qRT-PCR analyzed the enrichment of candidate miRNAs in RISCs in HepG2 and MHCC97H cells. c MiR-300 expression in five HCC cell lines and one human liver epithelium was evaluated via qRT-PCR. d qRT-PCR detected miR-300 expression in HepG2 and MHCC97H cells transfected miR-300 mimics or NC mimics. e YY1 mRNA and protein levels were measured by qRT-PCR and western blot in HepG2 and MHCC97H cells with the transfection of miR-300 mimics or NC mimics. f RNA pull down assay helped to determine the interaction of miR-300 with YY1. g StarBase predicted the binding sequence of miR-300 in YY1 3′UTR. h Luciferase reporter assay was performed to verify the interaction between miR-300 and YY1. i, j The effect of miR-300 mimics on the expression of factors related to WNT pathway was detected by qRT-PCR and western blot analyses. k TOP/FOP flash assay determined the activity of WNT pathway in cells transfected with NC mimics or mi-300 mimics. *P < 0.05, **P < 0.01