Fig. 3

SILAC quantitative proteomics characterizes MEST-regulated NF-κB pathway in lung cancer. A The scheme of two independent SILAC experiments, including forward labeling and reverse labeling. MEST-regulated proteins were identified by two independent SILAC experiments, and the differentially expressed proteins were analyzed by Ingenuity Pathway Analysis (B). MEST-driven functional signaling networks indicate involvement in the NF-κB signaling pathway. Red, upregulated proteins; green, downregulated proteins; white, speculative proteins. C MEST overexpression promotes nuclear translocation of NF-κB subunits (p65) in both A549 and H1299 cells. LaminB1 and GAPDH were used as markers for nucleus and cytoplasma, respectively. D A549 and H1299 overexpressing MEST were treated with either 2.5 μM of Bay11-7082 or DMSO, as indicated. Expression of NF-κB markers (including p-p65, p65, p-IκBα, and IκBα) was assessed by western blot analysis. Invasion and migration of treated cells were measured by transwell assay (E). F A549 and H1299 cells overexpressing MEST were treated with either 25 μM of MMP2 inhibitor I or DMSO; their invasion and migration abilities were determined by transwell assay. Scale bar, 100 μm. All data are representative of three independent experiments. Bars, S.D. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t-test)