Fig. 2: Characterization of the mitochondrial phenotype and metabolic features of heMSCs.

A Representative merged images of mitochondrial morphology and distribution abundance in heMSCs, hpMSCs, and A673 cells incubated with the mitochondrial potential-dependent dye Mito Red and the membrane-permeable live-cell labeling dye Calcein AM (in green), and shown by fluorescence microscopy at two different original magnifications. The pictures correspond to three lines of heMSCs and four lines of hpMSCs from different teratomas and individuals, respectively. Gray images, Mito Red pictures of the merged images above. Magnification bar: 10 μm. Observe the fragmented mitochondrial network in heMSCs and A673 cells. B Expression of metabolic enzymes in hpMSCs, heMSCs, and A673 cells, as determined by RT-qPCR, and referred to the mRNA of the mitochondrial translocon TIMM23. HK1 and GAPDH, glucolysis; ACLY, synthesis of cytosolic acetyl-CoA; Citrate synthase, Aconitase 2, IDH2, OGDH, and Fumarate hydratase, Krebs cycle; SHMT2, glycine synthesis; PCK2, conversion of oxaloacetate to phosphoenolpyruvate. Data obtained from one experiment performed in triplicate, and expressed as mean ± s.d. C Expression of the decoupling proteins UCP2 and UCP3 and glucose transporters GLUT1 and GLUT4 in hpMSCs, heMSCs, and Ewing sarcoma cells, and determined as in (B). In blue, Ewing sarcoma cell lines with the EWS::ERG (CHLA25 and COG-E-352) and EWS::FEV (TC205) fusions. D Levels of TCA metabolites in heMSCs, hpMSCs, and A673 cells. The statistics for comparisons of the heMSC and hpMSC groups in (B–D) were performed using the two-tailed unpaired t-test.