FIGURE 11.

C-R(7) containing peptides all reduce mitochondrial respiration, mitochondrial membrane potential, and generation of radical oxygen species in a dose-dependent matter. a, HEK293 cells were incubated with peptides (CN2097, C-R(7), R(7), n = 4 for each) at the indicated concentrations for 30 min at 37 °C. Mitochondrial respiration was analyzed in digitonin-permeabilized cells in measuring buffer (see “Materials and Methods”) in the presence of succinate and ADP using a computer-controlled closed micro-oxygen electrode chamber. Respiration rates were normalized to untreated controls and are presented as % change relative to control (n = 4). b, HEK293 cells were incubated with peptides CN2097, C-R(7), and R(7) (n = 4 for each) for 30 min at 37 °C, at the concentrations indicated. TMRM fluorescence was analyzed by flow cytometry. Results show that all three peptides resulted in a significant reduction in TMRM fluorescence, indicating a reduction of the mitochondrial membrane potential. c, detection of ROS in cultured HEK293 cells by the ROS-sensitive probe 2′,7′-dichlorodihydrofluorescein diacetate was assessed in the presence of 240 nm of either CN2097, C-R(7), or R(7) and compared with matched seeded and untreated cells. Results showed that all three peptides significantly reduced detectable ROS generation (n = 4 for each condition). *, significance from control p < 0.0001; p, error bars S.D.