Fig. 5

CircPLEKHM3 acts as a tumor suppressor in AKT1 and β-catenin signaling pathways. a Top five KEGG pathways enriched for differentially expressed genes upon knockdown of circPLEKHM3 in ovarian cancer cells. Differentially expressed genes were identified by RNA-seq data of circPLEKHM3 knockdown and scrambled control cells, and the 10 most significant genes are shown. b Immunoblotting analyses of P-AKT1, β-catenin, P-GSK3β, P21, and P27 in ovarian cancer cells with depletion of circPLEKHM3 or transfected with circPLEKHM3 overexpression and control vectors. c β-catenin signaling was activated in OV90 upon knockdown of circPLEKHM3. The pathway activation score was used to estimate the degree to which β-catenin signaling was activated in circPLEKHM3 knockdown and negative control (NC) OV90 cells. d Representative images (20× magnification) of immunohistochemical staining for P-AKT1 in immunodeficient mice injected with A2780 scramble and sh-circPLEKHM3 cells. e Immunoblotting analyses of AKT1 and P-AKT1 in circPLEKHM3 knockdown A2780 cells transfected with AKT1 siRNAs and negative controls (NC). f Cell growth and (g) migration assays were performed in circPLEKHM3 knockdown A2780 cells transfected with AKT1 siRNAs and negative controls. Migration assays were measured 24 h after transfection