Fig. 5

LncRNA-HGBC functions as a competing endogenous RNA by directly binding to miR-502-3p. a Luciferase activity in 293 T cells expressing lncRNA-HGBC binding miRNAs and empty luciferase reporter (left) or lncRNA-HGBC luciferase reporter (right). b Wild type and mutant lncRNA-HGBC sequences were cloned into pmirGLO vectors and co-transfected with miR-502-3p into 293 T cells followed by dual luciferase assay. c Anti-Ago2 RIP was used to pulldown endogenous RNAs associated with Ago2; IgG was served as the control. Ago2 in proteins from Ago2-RIP assay were measured by western blot. The levels of lncRNA-HGBC and miR-502-3p were measured by qRT–PCR and the data were presented as fold enrichment in Ago2 relative to input (*P < 0.05, ***P < 0.001). d MS2-RIP in NOZ cells followed by qRT-PCR to detect endogenous association between miR-502-3p and lncRNA-HGBC. miR-122 was a negative control. A schematic outline of the MS2-RIP strategy was shown (top). e qRT-PCR analysis of miR-502-3p expression in NOZ cells after knockdown of lncRNA-HGBC. f Transwell migration (top) and invasion (bottom) assays of GBC-SD cells that were transfected with the indicated plasmids. Scale bars, 200 μm. g After transfection of plasmids containing vector control, lncRNA-HGBC or lncRNA-HGBC-MUT, the proliferation of GBC-SD and EH-GB1 cells was measured using CCK-8 assays. *P < 0.05, ***P < 0.001