Figure 3 .

UBE2C regulates Akt phosphorylation and degradation

(A) The protein levels of Akt and p-Akt were analyzed by western blot analysis. (B) C2C12 cells were transfected with NC or si-UBE2C, followed by incubation with CHX. The cells were harvested at the designated time points for western blot analysis to evaluate Akt protein levels. (C) Greyscale scanning of the western blots was used to quantify the Akt protein levels. (D) C2C12 cells were transfected with siRNA and treated with SC79. The expression levels of MyoG, MyHC, Akt, and p-Akt were detected by western blot analysis. (E) C2C12 cells were transfected with plasmids to overexpress UBE2C and treated with LY294002. The expression levels of MyoG, MyHC, Akt, and p-Akt were analyzed by western blot analysis. (F,G) Immunofluorescence staining was performed on C2C12 cells treated as described in (D,E), with detection specifically targeting MyHC. Scale bar: 200 μm. Data are presented as the mean±SEM, n=3 per group. * P<0.05, and ** P<0.01. ns indicates no significant difference.