Skip to main content
. 2019 Apr 12;20(8):1810. doi: 10.3390/ijms20081810

Figure 6.

Figure 6

mir-590 enhanced β-catenin signaling. (A) A TOPFlash reporter assay was performed in SKOV3.ip1 cells stably transfected with mir-590 or its control empty vector (EV). mir-590 overexpression increased β-catenin transcriptional activity. Data represent the mean ± SEM (n = 3). (B) Cytosolic (Cyto) and nuclear (Nuc) fractions were prepared from EV and mir-590-overexpressing SKOV3.ip1 cells, and Western blotting was performed. mir-590 cells exhibited higher β-catenin in their nucleus. The densitometry readings of cytoplasmic and nuclear fractions were normalized to their corresponding GAPDH and Lamin B levels, respectively. (C) Immunofluorescence staining of β-catenin showed that SKOV3.ip1 cells stably overexpressing mir-590 had strong β-catenin signals in their nucleus. (D) Knockdown of CTNNB1 using siRNA reduced the mRNA and protein levels of β-catenin. (E) Silencing of CTNNB1 blocked the effect of mir-590 on spheroid formation in SKOV3.ip1 cells. Data represent the mean ± SEM (n = 10). Statistical analyses were performed using Student’s t-test (A and D), or two-way ANOVA/Student–Newman–Keuls test (E). *** p < 0.001, **** p < 0.0001.