Fig. 5.

Upregulation of OTUD6B-AS1 expression induced ccRCC cell apoptosis and altered cell cycle progression in vitro. a, b Flow cytometry assays were performed to analyze the cell cycle progression of the ACHN and OS-RC-2 cell lines transfected with plvx-OTUD6B-AS1 or the empty vector. The bar chart represents the percentages of cells in the G0/G1, S, or G2/M phase as indicated. All experiments were performed in biological triplicates with three technical replicates. c Western blot analysis was performed to assess the expression of CCNB1 and CCND1 in cells transfected with plvx-OTUD6B-AS1 or the empty vector. GAPDH protein was used as an internal control. d, e Flow cytometry was used to detect the apoptosis rates of the cells. LR, early apoptotic cells; UR, terminal apoptotic cells. f Western blot analysis was used to measure the levels of PARP, Bax and Bcl-2 in cells transfected with plvx-OTUD6B-AS1 or empty vector. β-Actin protein was used as an internal control. g Hoechst staining assay performed in ACHN and OS-RC-2 cells transfected with plvx-OTUD6B-AS1 or the empty vector. The nuclei of the apoptotic cells were dense or dyed (Cyan)