Fig. 5: SPOP controls pancreatic cancer phenotype partly through NANOG.

a Dynamic monitoring of the proliferation of SW1990 cells after NANOG downregulation in cells with SPOP knockdown using the iCELLigence RTCA Analyzer. NS, P > 0.05; ***P < 0.001. b The effects of NANOG down-regulation on the colony formation by SW1990 cells with SPOP knockdown. Quantification of the stained colonies is shown in the right panel. Data are the average of three experiments (mean ± SD). *P < 0.05; **P < 0.01; ***P < 0.001. c Cell cycle analysis of using flow cytometry after NANOG downregulation in SW1990 cells in which SPOP was knocked down. Quantification of cell percentage is presented on the right panel. NS, P > 0.05; **P < 0.01; ***P < 0.001. d Cell motilities were measured through testing the wound closure after NANOG downregulation in SW1990 cells in which SPOP was knocked down. Quantification of the wound closure is shown in the right panel. Data are the average of three experiments (mean ± SD). NS, P > 0.05; ***P < 0.001. e Transwell assays were used to detect the migration and invasion abilities after NANOG downregulation in SW1990 cells in which SPOP was knocked down. Scale bar = 100 μm. Quantification of the stained colonies is shown in the below panel. Data are the average of three experiments (mean ± SD). NS, P > 0.05; *P < 0.05; **P < 0.01; ***P < 0.001. f Western blot analysis of WCL derived from SW1990 infected with the indicated lentiviral shRNAs against SPOP and NANOG, and subjected to puromycin selection for 72 h before harvesting. g Representative western blot showing SPOP, NANOG, CDK2, and Snail expression in pancreatic cancer samples