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. 2015 Feb 23;17(9):1220–1230. doi: 10.1093/neuonc/nou369

Fig. 3.

Fig. 3.

Ras-mediated signaling modulates oxygen consumption rate (OCR) and spare respiratory capacity through inhibition of pyruvate dehydrogenase (PDH) activity. (A and B) The respiratory phenotype was defined in TR cells exposed to either vehicle control or the described agents 24 hours prior to analysis. To study mitochondrial function, cells were sequentially exposed to oligomycin (O), carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) (F), and rotenone/antimycin (R/A) in real time. The relative spare respiratory capacity (%) was calculated as (OCR following FCCP – baseline OCR)/baseline OCR. (C) PDH enzyme activity in the described cell lines and treatment conditions using a microplate assay kit. (D) Western blot was performed to determine levels of phosphorylated PDH (pPDH) in the described cell lines. Levels were quantified by determining the pPDH/PDH ratio for each line and then normalized to the ratio of T cells. (E) Western blot was performed to determine levels of pPDH in TR cells exposed to vehicle control or the stated agents. Levels were quantified by determining the pPDH/PDH ratio for each treatment condition and then normalizing to the ratio of vehicle control cells. *P < .05.