Fig. 2: Silencing TRIB3 inhibits the spheroid formation and tumor engraftment.

a GSEA demonstrating the enrichment of gene sets related to ES stem cell gene signatures in the ranked gene list of the top 10th percentile (n = 121) vs. the bottom 10th percentile (n = 121) of breast cancer patients expressing TRIB3 from the TCGA database (n = 1215). NES, normalized enrichment score. FDR, false discovery rate. b Flow cytometric analyses of representative unsorted MCF7 cells (left), isolated CD44^BrCD24^Di MCF7 cells (upper right), and isolated non-CD44^BrCD24^Di MCF7 cells (lower right). Contour plots (10% probability) depict the fluorescence of BCCs stained with fluorochrome-conjugated mAbs specific for human CD44 (y axis) or human CD24 (x axis). c, d Gene expression (c) by real-time PCR or immunoblots (d) of protein lysates of CD44^BrCD24^Di and non-CD44^BrCD24^Di MCF7 cells. The height of each column in the histogram indicates the fold increase in each gene or protein, as indicated on the left, of sorted CD44^BrCD24^Di MCF7 cells relative to that of sorted non-CD44^BrCD24^Di MCF7 cells. e, f Tumor spheroid formation of HMLE cells (e) transfected with control vector or TRIB3-expressing vector (as indicated at the top) or MCF7 cells (f) transfected with either CTRL-shRNA or TRIB3-shRNA. All data are shown as the number of mammospheres per 1000 cells. g, h Flow cytometric assays detecting the percentage of CD44^BrCD24^Di in HMLE cells transfected with a control vector or a TRIB3-expressing vector (g), or MCF7 cells transfected with either CTRL-shRNA or TRIB3-shRNA (h). i, j Tumor incidence in animals implanted with MCF7 cells (i) or MDA-MB-231 cells (j) transfected with either CTRL-shRNA or TRIB3-shRNA. Data are shown as the mean ± SEM; P > 0.05 was considered not significant (NS); *P < 0.05, **P < 0.01, and ***P < 0.001, compared with the vector or CTRL-shRNA group. Source data are provided as a Source Data file.