Figure 2

Ghrelin administration (for 4 weeks) normalizes altered gene expression profile in R6/2 mouse skeletal muscle. Normalized skeletal muscle (gastrocnemius) gene expression related to muscle contractility and atrophy (a) and mitochondrial biogenesis (b) at 14 weeks and after 4 weeks of ghrelin administration. Expression of Acta1 was significantly decreased, while the expression of Casp3, Casp8, Creb1, Smad3 and Traf-5 were significantly increased in R6/2 compared to WT mice. 4 weeks of ghrelin administration significantly normalized Casp8, Creb1 and Traf-5 expression (a). Normalized mitochondrial gene expression at 14 weeks illustrates significantly decreased expression of Mfn2, while the expression of Sirt1 was significantly increased. There was no significant effect of ghrelin administration (b). In panel c and d representative Western blot and densitometry of protein levels at 14 weeks are shown (c,d). We could see a trend for upregulation of TRAF-5 protein intensity (c) and activated CREB levels (d) in vehicle treated R6/2 mice compared to WT littermates. Ghrelin administration did marginally affect protein levels in R6/2 mice. Gene expression was analysed using the ΔΔCt method and normalized to Tbp and 18 S. Samples were processed in parallel on gels/blots, and stain-free method was used for normalization of protein levels. Full-length blots are presented in Supplementary Fig. 3. Data represent mean ± SEM of the following number of 8–11 animals/group. Statistical significance was determined by 2-way ANOVA with Bonferroni post hoc test for multiple comparisons. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001