Fig. 4

LncRNAs can crosstalk with miRNAs through direct binding. a Relative expression of lncRNAs in GC cells treated with mimics of miR-596 or miR-3620-3p were measured by real-time PCR. b Luciferase activity in GC cells co-transfected with luciferase reporter containing lncRNA-KRTAP5-AS1 and the mimics of miR-596, miR-3620-3p or mutant. Data are presented as the relative ratio of renilla luciferase activity and firefly luciferase activity. c Luciferase activity in GC cells co-transfected with luciferase reporter containing lncRNA-TUBB2A and the mimics of miR-3620-3p or mutant. Data are presented as the relative ratio of renilla luciferase activity and firefly luciferase activity. d The schematic diagram and real-time PCR results of the MS2-RIP method used to identify the binding between lncRNAs and miRNAs in both SGC-7901 and AGS cells. e The schematic diagram of the RNA pull down method used to identify the binding between lncRNAs and miRNAs in both SGC-7901 and AGS cells. GC cell lysates were incubated with biotin-labeled lncRNA-KRTAP5-AS1, lncRNA-TUBB2A, lncRNA-KRTAP5-AS1-mut, and lncRNA-TUBB2A-mut. MiRNA real-time PCR was performed after pull down process. Data are shown as mean ± s.d., n = 3. The data statistical significance is assessed by Student’s t-test. *P < 0.05, **P < 0.01