Ion is regulated by E2 in skeletal muscle in vivo to determine the effect of E2 on USP19 expression in gastrocnemius and soleus were female Mice Eierst the skirts removed. The ratios Of muscle mass to K Body weight tends to be green Be at M as it skirts mice in M mice Without Eierst Controlled Wrong, although the difference was not statistically significant. The treatment of mice M Without Eierst skirts with E2 replacement decreased the ratio Body weight ratio of muscle mass to K. The ratio Ratio of uterine weight on the K Body weight in ovariectomized M Mice was significantly decreased, and was won by replacing E2. In M Mice without Eierst skirts, USP19 decreased protein content in soleus and gastrocnemius and were restored by E2 replacement. Oophorectomy has entered Born USP19, the mRNA level decreased in the soleus, but not in the gastrocnemius muscle. E2 replacement increased Hte levels of USP19 mRNA in gastrocnemius and soleus. These results show that E2 regulates expression of USP19 in skeletal muscle in vivo. USP19 in myogenic differentiation E2 suppressed as a deubiquitinating enzyme to determine whether USP19 displaced in E2 Other appa myogenic differentiation is involved is involved, C2C12 myoblasts were transfected with USP19 siRNA, followed by the induction of differentiation. USP19 siRNA specifically struck the USP19 protein expression, but not the GAPDH protein in the presence or absence of E2, indicating the Dihydrofolate Reductase specificity con t and efficiency of USP19 We siRNAs. Removable USP19 obtained Ht the expression of MHC, tropomyosin and myogenin not only in cells but also in cells E2-treated vehicletreated. Indeed, USP19 knockdown prevents found Promotes myogenesis in the vehicle-treated cells and rescued the E2 formation of myotubes.
However, when C2C12 cells as exogenous wild-type USP19 w Overexpressed during differentiation, decreased expression of MHC and tropomyosin in the presence or absence of E2. In order to assess whether the inhibitory effect on myogenesis is required for USP19 deubiquitinating activity of t, mutated forms of activity T USP19 deubiquitinating missing were built, called USP19 and USP19. These mutants replace Ser and Ala for Cys at position 548 of USP19 are inactive. As shown in Fig. 5B, decreased the overexpression of wild-type USP19, the amounts of ubiquitin-labeled proteins, w While the mutant form of USP19 or USP19) dominant negative has backfired. Therefore, the effect of these mutants on the expression of protein markers were examined by myogenic differentiation. To reduce the F Ability, USP19 expression of MHC and tropomyosin, which has lost by the mutation in the presence or absence of E2. As n To search results, we examined the effect of E2 on the ubiquitylation of intracellular Other proteins in C2C12 cells. When C2C12 cells were grown in a differentiation in the presence ofMG132, protein levels were obtained Ht ubiquitin in a manner dependent Ngig of time. Exposure of C2C12 cells with suppressed levels of E2 protein ubiquitin. To determine the effect of the surcharge on the level of USP19 ubiquitin tagged proteins, C2C12 cells were transfected with USP19 siRNA followed, the culture in the presence or absence of E2. USP19 knockdown increased Hte ubiquitin in the presence or absence of E2.