Figure 6.

Phosphorylation of β-catenin at Ser311 facilitates its stability and translocalization into the nuclei in NSCLC. A, KEGG pathways functional analysis in invasive or non-invasive A549 cells expressing wild-type (WT), S311D (SD), or S311A (SA) of β-catenin. The invasive A549 cells expressing wild-type or mutants of β-catenin, transcriptome profiles were analyzed by microarray. The transcriptome data were clustered by gene probes and fold changes >1.5 indicated significant changes in cells expressing wild-type (WT), S311D (SD), or S311A (SA) of β-catenin. Gene Ontology (GO) and KEGG pathway analysis of transcriptome profiles was performed. The top 10 terms of KEGG pathways were analyzed, and the signaling pathways based on the number of signature genes in cells expressing invasive WT (left panel), SD (middle panel), or SA (right panel) compared to those expressing mock. ***p <0.001. B-C, A549 cells expressing RFP-tagged WT, S311D, or S311A of β-catenin were treated with 100 µg/ml cycloheximide in a time-dependent manner. B, Immunoblotting was performed to detect the levels of exogenous β-catenin using anti-β-catenin antibody. C, The relative levels of exogenous β-catenin were quantified using LI-COR Odyssey software (Li-COR Biosciences), normalized, and plotted. *p <0.05; **p <0.01; ***p <0.001. D, Immunofluorescence was performed with A549 cells expressing wild-type (WT), S311D β-catenin, S311A β-catenin, and β-catenin^mtGSK3β (mtGSK3β). RFP protein (red), α-tubulin (green), and DNA (DAPI, blue) are displayed. Scale bar, 20 µm. E-F, Quantification of the population of cells in the cytoplasm, nucleus, and both. n> 800.