Figure 1. K14Q MOTS-c has a reduced insulin sensitizing effect compared to WT MOTS-c in muscle and fat cell lines. (A) Differentiated C2C12 myotubes, were treated with WT and K14Q MOTS-c in the absence or presence of 10-nM insulin. The total and phosphorylated AKT (Ser473) levels were measured by MSD. (B) 3T3L1 Cells were incubated with 1-μg/mL insulin +/- WT MOTS-c or K14Q MOTS-c. At day 12, lipid droplets were measured by Nile Red staining. (C) Differentiated L6 myotubes were treated with WT and K14Q MOTS-c for 96hr. (D) Overexpression of WT MOTS-c, but not K14Q MOTS-c, increases glucose uptake in HEK293 Human embryonic kidney cells. HEK293 cells were transiently transfected with WT MOTS-c and K14Q MOTS-c for 72 hr. Glucose levels in the medium were measured. Glucose utilization was calculated by [Total amount of glucose in the media] – [remaining glucose in the media]. Data are reported as mean ± SEM of seven independent experiments. *** p < 0.001** p < 0.01, *p < 0.05.