Fig. 4

LIPH-4 acts as a sponge of miR-216b in ESCC cells. A Relative LIPH-4 expression levels in nuclear and cytosolic fractions of KYSE510 cells, measured by qPCR. U6 (nuclear retained) and GAPDH (exported to cytoplasm) were used as controls (n = 3). B Sequence alignment of miR-216b with binding sites in the wild-type (LIPH-4-wt) and mutant-type regions of LIPH-4 (LIPH-4-mut). C Relative luciferase activity in 293 T cells was assessed after co-transfection with the reporter plasmid (LIPH-4-wt or LIPH-4-mut) and miRNAs (miR-216b or NC mimics) (n = 3). D RIP assay was performed with an anti-Ago2 antibody. The levels of LIPH-4 and miR-216b were determined by qRT-PCR and presented as fold enrichment in Ago2 relative to IgG immunoprecipitates (n = 3). E Relative expression levels of miR-216b in ESCC and adjacent noncancerous tissue samples assessed by qRT-PCR (n = 53). F Negative correlation between miR-216b and LIPH-4. *P < 0.05, **P < 0.01, ***P < 0.001