Research Paper Volume 13, Issue 7 pp 10749—10769

Figure 6. MiRNA-150 induces Hh and Wnt signaling activation and EMT via SUFU. (A) MiRNA-150 inhibition decreased β-catenin-dependent transcriptional activity, and this decrease was reversed by SUFU siRNAs. A TOPFlash/FOPFlash assay was conducted in BGC-823 cells transfected with miRNA-150 inhibitors or co-transfected with 150-inh and SUFU siRNAs. (B, C) Interference of SUFU alleviated the β-catenin cytoplasmic translocation induced by miRNA-150 inhibition. (B) β-catenin expression was detected in the cytoplasm and nucleus. (C) Confocal immunostaining of β-catenin was performed to visualize the cytoplasmic or nuclear localization of β-catenin. (D, E) MiRNA-150 inhibitors reduced the expression of Hh signaling genes and EMT markers, and SUFU siRNA rescued this inhibitory effect. The expression of Hh signaling and EMT proteins was detected in BGC-823 cells by western blotting after transfection with miRNA-150 inhibitors or co-transfection with 150-inh and SUFU siRNAs. (F) The mRNA levels of some Hh-, Wnt- and EMT-related genes were reduced under miRNA-150 inhibition, but increased somewhat after SUFU gene knockdown. *p<.05.