Figure 4 from Multistate Gene Cluster Switches Determine the Adaptive Mitochondrial and Metabolic Landscape of Breast Cancer

Mitochondrial function: MB1_UF cells show active functional biogenesis in contrast to the MB1_LF phenotype. A, Two-dimensional distribution (PC1 vs. ranking index, shown for the MB1 bicluster switch) of biopsies obtained for generating PDX samples from the study by Bruna and colleagues (38), overlaid in left panel with Pam50 categories and in the right panel with PDX status indicating whether the sample has grown from the biopsy to PDX status. B, Cell line scoring to sort cell lines to the MB1 switches. Volcano plot of the scores and −log₁₀ of the P values are shown. The cell lines chosen for experimental work are highlighted in red. C, OCR in cell lines grouped by their switch state (MB1_UF/MB1_LF). Reserve, ATP synthesis coupled, and uncoupled respirations were quantified using FCCP and oligomycin treatments. Two-way ANOVA with Sidak multiple comparisons test. D, Average cellular TMRM intensity grouped by cell line switch state (MB1_UF/MB1_LF). Data were obtained from high content microscopy from >1K cells per group. Unpaired Student t test. E, Relative expression of respiratory chain subunits quantified from Western blot analysis. Results are grouped by switch state (MB1_UF/MB1_LF). Two-way ANOVA with Sidak multiple comparisons test. F, High content imaging analysis of mitochondrial biogenesis, structure, and function in MB1_UF and MB1_LF cells. Scale bars, 10 μm. Quantifications of the picoGreen integrated intensity (readout for mtDNA content), mitochondrial number, total volume, and individual mitochondrial volume are shown. Unpaired Student t tests. ns, nonsignificant, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.