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. Author manuscript; available in PMC: 2013 Jun 11.
Published in final edited form as: Nat Commun. 2012;3:1271. doi: 10.1038/ncomms2236

Fig. 3.

Fig. 3

MtErbB2 reprograms cellular metabolism from oxidative phosphorylation toward glycolysis. (A) Mitochondrial and whole cell lysate of 231V, 231ErbB2WT, 231ErbB2Mito, and 231ErbB2ΔMTS cells were isolated and analyzed by Western blotting. Protein amounts of each fraction loaded: mitochondria: 10 ug; whole cell lysate: 30 ug. Integrin β1 was used as plasma membrane maker and loading control; mtHSP70 was a mitochondrial maker and loading control; α-Tubulin was a cytoplasm maker; KDEL was an ER marker; EEA1 was an early endosomes marker; Golgi Complex was a marker for the detection of Golgi; LAMP2 was an lysosome marker and Lamin B1 was a nucleus marker. (B) Oxygen consumption rates. Oxygen consumption rates of 231V, 231ErbB2WT, 231ErbB2Mito, 231ErbB2ΔMTS and 231ErbB2MitoKM cells were measured. The oxygen consumption rate was calculated on the basis of the maximal rate of change in relative fluorescence units (DFU/second). (C) Activities of the mitochondrial electron transport chain complexes in 231V, 231ErbB2WT, 231ErbB2Mito, 231ErbB2ΔMTS and 231ErbB2MitoKM cells. Activities are presented as milliunits of O.D. value per min and were normalized by the amounts of the mitochondrial proteins. (D) Mitochondrial membrane potential (ΔΨm) of 231V, 231ErbB2WT, 231ErbB2Mito, 231ErbB2ΔMTS and 231ErbB2MitoKM cells was detected using JC-1 staining. The aggregate form of JC-1 staining represents healthy mitochondria. (E) The cellular ATP/ADP ratio of 231V, 231ErbB2WT, 231ErbB2Mito, 231ErbB2ΔMTS and 231ErbB2MitoKM cells were detected. (F) MtErbB2 increased glucose uptake. 231V, 231ErbB2WT, 231ErbB2Mito, 231ErbB2ΔMTS and 231ErbB2MitoKM cells were cultured in medium containing 10% fetal bovine serum (FBS) and the glucose uptake were measured. Data are shown as a percentage relative to MDA-MB-231V. (G) MtErbB2 increased lactate production. 231V, 231ErbB2WT, 231ErbB2Mito, 231ErbB2ΔMTS and 231ErbB2MitoKM cells were cultured in medium containing 10% FBS and the lactate production in the medium was measured. Data are shown as a percentage relative to 231V. Columns, mean of three independent experiments; bars, SE. *, P<0.05, **, P<0.01, ***, P < 0.001.