Figure 6 from Multistate Gene Cluster Switches Determine the Adaptive Mitochondrial and Metabolic Landscape of Breast Cancer

Protein expression profiles match transcriptomic switches. A, Western blot analysis of 29 metabolic enzymes across five cell lines representing MB1_UF and MB1_LF switch positions and cell states. Representative blots from n ≥ 3 independent experiments, used for quantification of signals, normalizing density values to either grp75 or β-actin. B, D, and E, Clustering and quantitative analysis of enzyme expression levels obtained from A. Values were separately normalized to either mitochondrial (left) or cytosolic (right) markers, grp75 or β-actin. The comparison of mitochondrial abundance based on the grp75/β-actin ratio is shown at the bottom in B. Heatmaps are based on a pairwise, robust to outliers distance measure of the mean protein expression values. The k-means clustering. D and E, Two-way ANOVA with uncorrected Fisher LSD multiple comparison tests. C, Heatmap and k-means clustering of transcript CV values of the 29 enzymes from the METABRIC dataset (see Figs. 1 and 2, positive values predicting higher expression of genes in MB1_UF and negative values corresponding to higher expression in MB1_LF) and scaled protein expression levels (wCV) averaged across all cell lines, normalized to grp75 (wCV_mito) or β-actin (wCV_cyto). ns, nonsignificant, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001).