Figure 2

Palmitate activates ROS and signalling pathways in muscle cells. L6 muscle cells were treated for 24h with 0.2 mM palmitate (PA), palmitoleate (PO) or BSA control. A) Reactive oxygen species were analyzed using DCFDA, normalized to the protein content and expressed as percent of the BSA control. Results are mean ±SEM (n=4) analyzed using 1-way ANOVA, **p<0.01 vs BSA control. B-E) Proteins were extracted with standard lysis buffer and analyzed by western blotting using specific antibodies to IκBα and the phosphorylated forms of ERK, JNK and p38 MAPK. Results were normalized to the loading control actinin-1, expressed relative to the BSA control as mean ±SEM (n=4) and analyzed using 1-way ANOVA, **p<0.01, ***p<0.001 vs BSA control. F) Representative blots for B-E. G) Time-course of IκBα degradation. The changes of IκBα were calculated to the BSA control as indicated by the dotted line set at 1.0 from the y-axis **p<0.01 vs BSA control.