Fig. 4

MRVI1-AS1 recruits CELF2 to stabilize SKA1 mRNA in HCC cells. A Data from the public datasets StarBase (http://starbase.sysu.edu.cn) showed that CELF2 was the potential RNA-binding protein for both MRVI1-AS1 and SKA1 mRNA. B Public platform RPIseq (http://pridb.gdcb.iastate.edu/RPISeq) was used to predict the interaction probability between CELF2 protein and MRVI1-AS1 or SKA1 mRNA (the interaction probabilities > 0.5 means positive result. RF random forest, SVM support vector machines). C RNA pull-down assay was performed by using biotin-labeled MRVI1-AS1 and the antisense-MRVI1-AS1 to assess the interaction between CELF2 protein and MRVI1-AS1. D RIP assay using antibody against CELF2 was used to explore the interaction between lncRNA MRVI1-AS1 and CELF2 protein in MHCC-97H and Hep3B cells. IgG served as the control. Then, immunoprecipitated RNA was purified and analyzed by RT-qPCR (mean ± SD; n = 3). **P<0.01, **P<0.001, Student’s t test. D–F RIP assay using antibody against CELF2 was used to explore the interaction between lncRNA MRVI1-AS1 and CELF2 protein in MRVI1-AS1 knockdown or MRVI1-AS1-overexpressing subclones of MHCC-97H and Hep3B cells. IgG served as the control. Then, immunoprecipitated RNA was purified and analyzed by RT-qPCR (mean ± SD; n = 3). **P<0.01, **P<0.001, two-way ANOVA. G, H Western blot and RT-qPCR analysis were applied to determine the regulatory relationships among MRVI1-AS1, CELF2, and SKA1 mRNA (mean ± SD; n = 3). **P < 0.001, **P < 0.001, two-way ANOVA