TRIM27 and SIX3 expression levels and correlation analysis in lung cancer cells and tissues. (A) A549 cell lysates underwent co-immunoprecipitation with anti-SIX3 or control IgG antibody. The immunoprecipitates were then immunoblotted with the indicated antibodies. TRIM27, SMURF2, and SIX3 expression in lung cancer tissues (n = 138) and adjacent normal lung tissues (n = 30) in our independent hospital cohort was detected by quantitative real-time PCR (B) and immunohistochemistry (D). (C, E) Correlation analysis of SIX3, SMURF2, and TRIM27 in lung cancer tissues (n = 138). Statistical analyses were performed using the Chi-square test. (F) TRIM27 and SIX3 expression in cells from the NSCLC cell lines H1975, A549, and H358, and the human bronchial epithelial cell line 16HBE were measured by western blot analysis. Survival probability of patients with lung cancer from a Kaplan-Meier Plotter database (G) and our independent hospital cohort (H, I). Scale bar: 50 μm. All experiments were repeated at least three times, and data are represented as mean ± SD. (B) ***P < 0.001 (Mann-Whitney U test). (F) *P < 0.05, ***P < 0.001 (one-way ANOVA followed by Dunnett’s test).