Fig. 4

EVI5 affects the proliferation of NSCLC cells by regulating the accumulation of Emi1. a Western blot analysis of EVI5 and Emi1 protein levels in 20 randomly selected NSCLC tissues and paired noncancerous lung tissues. The band density ratios represent the relative expression levels of EVI5 and Emi1. b Relative protein expression levels of EVI5 and Emi1 in 20 paired NSCLC tissues. The Y axis indicates the log10 transformed fold change in the T/N protein expression ratios of EVI5 and Emi1. The number of each specimen is indicated below the X axis. c Correlation between EVI5 and Emi1 protein expression in 20 paired NSCLC tissues. The X and Y axes indicate the log10 transformed fold change in the T/N protein expression ratios of EVI5 and Emi1, respectively (P < 0.001). d Co-immunoprecipitation of EVI5 and Emi1 are shown. Protein were immunoprecipitated and detected from lysates of control and EVI5-overexpressing or EVI5-KO A549 cells using a specific monoclonal antibody. e The levels of Emi1 and downstream signaling molecules were measured; the levels of Emi1, p-Akt, p-Erk, CyclinA2, and CyclinD1 were significantly decreased in EVI5-knockdown cells compared with control cells. f CCK-8 assay to assess the viability of EVI5-overexpressing A549 cells transfected with siRNAs targeting Emi1 or with si-NC. Cell viability was assessed at 24, 48 and 72 h. g Representative images of the clonogenic assay results for cell proliferation in EVI5-overexpressing A549 cells transfected with siRNAs targeting Emi1 or with si-NC. Bar charts showing the clonogenic growth of cells. h EVI5-overexpressing A549 cells were transfected with siRNAs targeting Emi1 or with si-NC, and the expression of various proteins was then measured by western blot analysis. β-actin was used as the internal control. Bars represent mean ± SD from three independent experiments. Significant differences compared with the control: ** P < 0.01; ***P < 0.001. Abbreviations: PLVX, PLVX-IRES-Neo vector