TWEAK induces invasion, NIK mRNA expression, and noncanonical NF-κB nuclear translocation. (A) Quantification of BT116 invasion density at 48 h with or without 10 ng/ml TWEAK in the collagen matrix. Data represent average numbers of invading cells per 1-mm2 field (n = 3 wells) ± S.E.M. *p < 0.05 relative to vehicle control using unpaired Student’s t-test. (B) Western blot analysis of proteins from cytoplasmic and nuclear fractions of BT116 cells stimulated with 10 ng/ml TWEAK for the indicated times using the indicated antibodies. (C) Western blot analysis of cytoplasmic and nuclear fractions as in Figure 3B post-stimulation with 10 ng/ml TNFα for the indicated times. (D) qRT-PCR analysis of NIK mRNA expression from BT116 cells. mRNA was collected at indicated time points after stimulation with TWEAK (10 ng/ml) or TNFα (10 ng/ml). Data represent average mRNA expression relative to RPLP0 mRNA (n = 3 replicates) ± S.D.