Fig. 1
The nuclear localization of NF-κB2 (p52) and RelB was facilitated in LM05 cells via NIK upregulation at the protein level. A Schematic representation of the in vivo selection process using an orthotopic xenograft (OX) and tail vein injection (TVI). Luciferase-expressing MDA-MB-231 (parental cells) cells were transplanted into NOD-SCID mice by OX or TVI. Subsequently, lung metastatic cells were collected and established from lung tissue with metastases. These cells were reinjected into NOD-SCID mice using the same xenograft model to concentrate these cells with higher lung metastatic activity. B Cell growth curves of Parent, LM05 and LM1-2-1 cell lines on planar culture (n = 3, two-way ANOVA followed by Tukey’s multiple comparison test). C Representative tumor growth curves (lower, n = 4 per group, two-way ANOVA followed by Tukey’s multiple comparison test) of NOD-SCID mice orthotopically injected with parental, LM05 or LM1-2-1 cells at 0 week and 5 weeks. D Integrative signaling analysis of LM05 and LM1-2-1 cells was analyzed by microarray expression data. The microarray expression results were shown as a heatmap of the log2-fold change in each cell compared with the parental cells. E. Representative primary blots and heatmap of integrative signaling analysis by western blotting. The protein production data of western blotting were shown as a heatmap of the log2-fold change in each cell compared with the parental cells. F Western blotting analysis of NF-κB1 (p50), NF-κB2 (p100/52), RelA, RelB, α-Tubulin and Histone H3 in whole cells and the nuclear and cytoplasmic extracts of the parental, LM05, and LM1-2-1 cells. G Western blotting (upper) and qRT-PCR (lower, n = 3, one-way ANOVA followed by Tukey’s multiple comparison test) results of NIK protein and mRNA in the parental, LM05, and LM1-2-1 cells. For western blotting, all cell lines were either untreated or treated with MG132 (10 μM for 4 h). All data are representative of three independent experiments and are shown as the mean ± SEM. n.s. not significant. * P < 0.05