Fig. 6. Effect of palmitate and/or unsaturated fatty acids on mitochondrial ROS production. L6 myotubes were incubated for 16 h in media containing D-glucose (5 mM) supplemented with or without palmitate (PA, 0.4 mM) or unsaturated fatty acids palmitoleate (PO, C16:1), Oleate (OL, C18:1) or linoleate (LO, C18:2) at the concentrations indicated prior to analysis of (A) superoxide production (five measurements/condition/experiment); (B and C) H₂O₂ production and (D) mitochondrial membrane potential using spectral analysis to monitor JC-10 aggregate: monomer content. Data are presented as mean + SEM from a minimum of four separate experiments. The asterisk indicates a significant change (P<0.05) to the control condition (lacking any fatty acid additions) or between the indicated bars.