Fig. 3: miR-200 FKO increases lysosomal content and induces an aberrant metabolic phenotype.

A SA-β-Gal staining for A-NTC cells, the miR-200 FKO clone A-31, and A-26 with residual miR-200s expression in vitro. Scale bar, 50 μm. B The percentage of cells staining positive for SA-β-Gal from A. C SA-β-Gal staining for sections from cell-derived xenografic tumor-bearing these three clones in vivo. Scale bar, 100 μm. D The percentage of cells staining positive for SA-β-Gal from C. E Oxygen consumption rate (OCR) was measured using the Seahorse analyzer in these clones with the treatment of oligomycin, FCCP, and a mix of antimycin A and rotenone. F Basal OCR and G Spare respiratory capacity (SRC) was determined from E. H Cellular ROS was measured using flow cytometric analysis for DCF stained cells. I Western blot analysis of mitochondrial protein complexes in these clones. J Extracellular acidification rate (ECAR) was measured using the Seahorse analyzer in these clones with the treatment of glucose, oligomycin, and 2-deoxy-glucose (2-DG). K ECAR and L Glycolysis reserve was determined from J. M Lactate production was measured in the culture medium of these cells. N Western blot analysis of glycolysis-related proteins in these clones. O Staining (left) and quantification of mitochondrial activity (right) of these clones with Mito-Tracker (red). Scale bar, 20 μm. Data represent the mean ± SEM of triplicate independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001. P Representative images of transmission electron microscopy for observing the morphology and structure of mitochondria from these clones. Scale bar, 5 μm.