Extended Data Fig. 10. Phenotypic effects of BACH1 correlate significantly with downstream target gene expression.
(a) Top 20 cell-motility-related biological processes from gene set enrichment (GSE) analysis performed for genes positively correlated with BACH1 (left) and for genes negatively correlated with RKIP (right) in the TCGA BRCA set (breast cancer; provisional, n = 1100), ranked by fold enrichment coefficient. Cyan-labeled processes are shared between the two GSE sets. (b) Top 10 genes with expression positively correlated with BACH1 expression (left) and negatively correlated with RKIP expression (right) in the TCGA BRCA set. Cyan-labeled genes are shared between two lists. (c) Relative BACH1 mRNA expression z-score with respect to the average expression of diploid samples in the TCGA BRCA Provisional dataset (n = 1100). Pearson Correlation value between total BACH1 and RKIP was r = −0.3690, P < 0.0001; while for BACH1 lower and higher expression groups were r = −0.3861, P < 0.0001 and r = 0.0062, P = 0.8930 respectively. (d) Correlation between RKIP mRNA and corresponding BACH1 mRNA level at each Dox concentration in both low-noise mNF-BACH1 (BL) and high-noise MB231 mNF-BACH1 (BH) clones, r = −0.2486, P = 0.4359. Samples were averaged with n = 3 technical replicates and normalized to the mean of corresponding uninduced sample, Pearson Correlation. (e) The microRNA-mediated BACH1 knockdown, hemin-mediated BACH1 degradation and RKIP overexpression repress invasiveness comparably. Two-tailed t-test between control samples and every other condition, n = 3, *P < 0.05, **P < 0.01. (f) mRNA level changes of HMOX1, MMP1 and CXCR4 for increasing Dox concentrations in high-noise mNF-BACH1 (BH) MB231 clones with respect to the corresponding uninduced sample. n = 3; one-way ANOVA with Dunnett’s multiple comparisons test between each dose and uninduced controls, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (g) Dose-response of CCND1 expression correlates significantly with monotone reduction of proliferation as BACH1 levels increase. r = −0.8988, P = 0.0381, n = 3, Pearson Correlation.