Fig. 6
CircMYO10 inhibits miR-370-3p to upregulate RUVBL1 expression and promotes the interaction between RUVBL1 and β-catenin/LEF1 complex and thus activates Wnt/β-catenin signaling. a MG63 cells were stably transfected with plasmids indicated while TOP/FOPFLASH plasmids, together with Renilla luciferase plasmids, were transiently transfected to measure the luciferase activity. Renilla luciferase activity was used as an internal control. Data represents the mean ± SD (n = 9). b-c MG63 cells were stably transfected with expression vectors as indicated. Pre-miR-370-3p reduced the expression of RUVBL1 and β-catenin in MG63 cells. Lysates were incubated with either anti-β-catenin or anti-LEF1. Immunoprecipitated complexes were subjected to western blot analysis. Anti-RUVBL1 antibodies derived from different species were used to avoid the detection of heavy chains. d Western blot analysis of RUVBL1, β-catenin, C-myc and CyclinD1 in MG63 cells and U2OS cells stably transfected expression vectors as indicated. e Transfection with miR-370-3p sponge promoted the migration and invasion ability of both MG63 and U2OS cells which was partially abrogated by either ShRUVBL1 or TIP49D302N. Data represents the mean ± SD (n = 3). (f) TCF/LEF luciferase activity assays were conducted in MG63 cells transfected with expression vectors indicated. Data represents the mean ± SD (n = 9). g-h ShcircMYO10 downregulated the expression of g β-catenin, g RUVBL1 and h LEF1, which was restored by co-transfection with either RUVBL1 or miR-370-3p sponge in MG63 cells. Proteins immunoprecipitated with β-catenin either LEF1 were subjected to western blot analysis. i ShcircMYO10 decreases the expression of RUVBL1, β-catenin, cyclinD1, C-myc, N-cadherin and vimentin with E-cadherin upregulated, and the change was partially attenuated by co-transfection with either miR-370-3p sponge or RUVBL1. j The enhanced migration and invasion ability of MG63 and U2OS cells were partially abrogated by co-transfection with any of Pre-miR-370-3p or shRUVBL1 or TIP49D302N. Data represents the mean ± SD (n = 3). Three independent assays were performed in the above assays. a, e, f, j * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t-test)