Impairment of mitochondrial respiratory function as an early biomarker of apoptosis induced by growth factor removal

Abstract

Intracellular signaling pathways not only control cell proliferation and survival, but also regulate the provision of cellular energy and building blocks through mitochondrial and nonmitochondrial metabolism. Wild-type and oncogenic RAF kinases have been shown to prevent apoptosis following the removal of interleukin 3 (IL-3) from mouse pro-myeloid 32D cells by reducing mitochondrial reactive oxygen species production. To study primary effects of RAF on mitochondrial energy metabolism, we applied high-resolution respirometry after short-term IL-3 deprivation (8 h

Sex-dependent effects of developmental hypoxia on cardiac mitochondria from adult murine offspring

“…When analyzing the effect of sodium azide on H202 production, H202 was expressed as a percentage of the amount of O2 consumed (H202/O2). To estimate mitochondrial efficiency of ATP production, the OXPHOS-coupling efficiency ratio was calculated as 1 -(LEAKCI / OXPHOSCI), and the respiratory control ratio (RCR) was calculated as OXPHOSCI / LEAKCI) 33 .…”

Mitochondrial metabolic study guided by proteomics analysis in hepatocellular carcinoma cells surviving long-term incubation with the highest dose of sorafenib

“…After a long-term incubation, the LI72h and HI72h HepG2 cells were carefully washed twice in PBS and harvested via trypsin digestion; the cells were then resuspended in MEM culture medium at a cell density of 5×10 5 cells•ml -1 in 2-ml chambers stirred at a speed of 750 rpm. The cellular mitochondrial respiratory rate was measured at 37 °C with a high-resolution mitochondrial respirometer (Oxygraph-2k, Oroboros Instruments, Innsbruck, Austria) [49]; real-time data acquisition of O2 concentration (nmol•ml -1 ) and O2 flow per 10 6 cells (pmol•s -1 •10 -6 cells) were performed using the DataLab software (Oroboros Instruments, Innsbruck, Austria). Routine respiration was measured when the respiration was stabilized.…”

In Vitro Assays for the Assessment of Impaired Mitochondrial Bioenergetics in Equine Atypical Myopathy

“…In all protocols, digitonin (Dig; 0.01 mM) was added before ADP. Optimal digitonin concentration was determined by careful titration experiments as previously described [ 29 ]. In all SUIT protocols, ADP-stimulated respiration represents OXPHOS capacity, P whereas ET capacity, E was obtained by addition of the uncoupler FCCP, rendering this state as not limited by the capacity of the phosphorylation system [ 28 ].…”