Figure 4

miR-130a activates YAP through repression of VGLL4. (A) miR-130a activates YAP transcriptional activity. CTGF-luciferase reporter was co-transfected with other plasmids into HEK293T cells as indicated for luciferase assay. Experiments were performed in duplicates. (B) miR-130a inhibition suppresses YAP transcriptional activity in a VGLL4-dependent manner. HEK293T cells stably expressing CTGF-luciferase reporter were transfected as indicated for luciferase assay. Experiments were performed in duplicates. ^* indicates a P-value of < 0.05 as calculated by student's t-test. (C) miR-130a promotes YAP target gene expression, which is blocked by VGLL4. Control or VGLL4-overexpressing HepG2 cells were transfected with NC or miR-130a mimic for gene expression analysis. Experiments were performed in triplicates. (D) miR-130a promotes Hippo pathway target gene expression in a YAP-and-TAZ-dependent manner. HepG2 cells were transfected as indicated for gene expression assay. Experiments were performed in triplicates. (E) miR-130a inhibition suppresses YAP target gene expression in a VGLL4-dependent manner. HMLE cells were transfected and gene expression was determined. Experiments were performed in triplicates. (F) miR-130a regulates YAP binding to target gene promoter. ChIP was performed using control IgG or antibodies against TEAD1 and YAP on HepG2 stable cell lysates. Relative enrichment of CTGF promoter was determined by quantitative PCR in triplicates. (G) miR-130a regulates YAP-TEAD1 interaction. HepG2 cells stably expressing miR-130a precursor or sponge and respective empty vectors were lysed and immunoprecipitated with anti-YAP or anti-TEAD1 antibodies. Lysates and immunoprecipitates were analyzed by western blots.