Fig. 5

The interaction of FOXM1 with STIL is required for STIL-induced tumorigenic abilities. a HEK293T cells were transiently co-transfected HA-FOXM1 with full-length or various truncated Flag-STIL mutants including STIL-N (a.a. 1–628), STIL-M (a.a. 420–780) and STIL-C (a.a. 780–1288) as indicated. Protein complexes were immunoprecipitated using anti-Flag antibody and analyzed by Western blotting using indicated antibodies. b HEK293T cells were transiently co-transfected HA-FOXM1 with the full length or various truncated GFP-STIL mutants (a.a. 1–1061, a.a. 437–1288, and the deleted coiled-coil domain (ΔCC)). Protein complexes were immunoprecipitated using anti-HA antibody and analyzed by Western blotting. c, d NCI-H1299 cells were transiently transfected with GFP-STIL-WT or GFP-STIL mutant (ΔCC), and the expressed proteins were analyzed by Western blotting (c, upper panel). Cell migration and invasion abilities were analyzed by Boyden chamber assay (c, lower panel), while the SLUG promoter activity was measured by reporter assay (d, upper panel) and the SLUG mRNA level (d, lower panel) was measured by qPCR method. Data information: Statistical data in c, d represent the mean ± SD (n = 3 independent experiments). Significance is determined by t-test (NS, not significant; **p < 0.01; ***p < 0.001)