Fig. 3

STIL activates the EMT signaling pathway to promote cancer cell migration and invasion. a The major biological functions analyzed by GSEA analysis of the transcriptome differences between CL1-0 cells and CL1-0 cells overexpressing GFP-STIL under Dox treatment for 48 h. The red bar indicates the most significant biological function. b The relative mRNA levels of EMT-TFs (SLUG, SNAI1, and TWIST), the EMT-regulator MMP9, and the CS marker CD44, as measured by qPCR method in Dox inducible CL1-0 cells overexpressing GFP-STIL (left panel) and in STIL-knockdown CL1-5 cells (right panel). c The protein levels of EMT regulators were analyzed by Western blotting in Dox inducible CL1-0 cells overexpressing GFP-STIL (upper panel) and STIL-knockdown CL1-5 cells (lower panel). Tubulin served as loading control. d SLUG promoter activity was measured by reporter assay in HEK293T cells transiently transfected with a pGL3/SLUG promoter-luciferase plasmid and the indicated constructs. e STIL protein levels in cytoplasmic (C) and nuclear (N) fractions analyzed by Western blotting in the indicated cells. Lamin A/C and tubulin were used as nuclear and cytoplasmic markers, respectively. The percentage of subcellular distribution is also shown. f STIL protein levels derived from the subcellular fractions of cytosol, membrane, nuclear-soluble, chromatin-bound, and cytoskeleton in NCI-H1299 cells were analyzed by Western blotting. Tubulin, EGFR, lamin A/C, histone H3 and vimentin were used as cytoplasmic, membrane, nuclear-soluble, chromatin-bound, and cytoskeletal markers, respectively. Data information: Statistical data in b and d represent the mean ± SD (n = 3 independent experiments). Significance is determined by t-test (NS, not significant; **p < 0.01; ***p < 0.001)