Figure 7.

PHB2 promotes tumorigenesis by regulating RACK1 and RACK1-associated proteins and downstream signaling in vitro. (A) Endogenous Co-IP assays between RACK1 and integrin β1 using an anti-RACK1 antibody in A549 cells with stable PHB2 overexpression. (B)-(D) Representative immunoblots for PHB2, p-Akt, Akt, p-FAK, FAK, and β-actin and their densitometric quantification in A549 cells with stable PHB2 overexpression. (E)-(H) Representative immunoblots for PHB2, RACK1, p-Akt, Akt, p-FAK, FAK, and β-actin and their densitometric quantification in A549 cells with stable PHB2 overexpression transfected with a specific siRNA targeting RACK1. (I) Time-dependent cell viability of A549 cells from the indicated groups measured by CCK-8 assay. (J)-(K) Representative images and quantitation of colony formation of A549 cells from the indicated groups. These tests were repeated three times independently. (L) Representative images of IHC staining for RACK1 in 48 paired NSCLC tumor and adjacent normal tissues. (M) Quantitative analysis of RACK1 protein expression in tumor and tumor-adjacent tissues. (N) Correlation between the protein levels of RACK1 and PHB2 in 48 NSCLC tissues calculated using Pearson's correlation test (r = 0.3117, P < 0.05, n = 48). Data are presented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. Abbreviations: IHC, immunohistochemistry; siRNA, small interfering RNA.