Journal: Nutrition & Metabolism

Article Title: Supplementation of carnitine leads to an activation of the IGF-1/PI3K/Akt signalling pathway and down regulates the E3 ligase MuRF1 in skeletal muscle of rats

doi: 10.1186/1743-7075-10-28

Figure Lengend Snippet: Relative protein concentrations of total and phosphorylated mTOR at Ser 2448 and Ser 2481 and calculated phospho - mTOR (Ser 2448 )/total mTOR and phosphor - mTOR (Ser 2481 )/total mTOR ratios in M. quadriceps femoris of rats fed either a control diet (0 mg carnitine/ kg diet; Control) or a diet supplemented with 1250 mg carnitine/ kg diet (Carnitine) . ( A ) Representative immunoblots specific to total mTOR, phosphor-mTOR (Ser 2448 ), phosphor-mTOR (Ser 2481 ) and α-Tubulin as internal control are shown for one animal per group; immunoblots for the other animals revealed similar results. ( B ) Bars represent data from densitometric analysis and represent means ± SD (n = 6/group); bars are expressed relative to the protein level of the control group (= 1.00). * indicates a significant difference to the control group (P < 0.05).

Article Snippet: After incubation the membranes overnight at 4°C in blocking solution (5% nonfat dried milk powder), membranes were incubated with primary antibodies against MuRF1 (polyclonal anti-MuRF1 antibody; Santa Cruz Biotechnology, Inc., Santa Cruz, Ca, USA), atrogin-1/Fbx32 (polyclonal anti-Fbx32 antibody; Abcam, Cambridge, UK), ubiquitin (polyclonal anti-ubiquitin antibody, Abcam, Cambridge, UK), Akt1 (polyclonal anti-Akt1 antibody; Cell Signaling Technology, Boston, MA, USA), hosphor-Akt1 (at Ser 473 ) (monoclonal anti-pAkt1 antibody; Cell Signaling Technology, Boston, MA, USA), FoxO1 (polyclonal anti-FoxO1 antibody; Cell Signaling Technology, Boston, MA, USA), hosphor-FoxO1 (at Ser 256 ) (polyclonal anti-pFoxO1 antibody; Cell Signaling Technology, Boston, MA, USA), mTOR (polyclonal anti-mTOR antibody; Cell Signaling Technology, Boston, MA, USA), hosphor-mTOR (at Ser 2481 ) (polyclonal anti-pmTOR antibody; Cell Signaling Technology, Boston, MA, USA), hosphor-mTOR (at Ser 2448 ) (polyclonal anti-pmTOR antibody; Cell Signaling Technology, Boston, MA, USA) and α-Tubulin (polyclonal anti-α-Tubulin antibody; Cell Signaling Technology, Boston, MA, USA) and GAPDH (monoclonal anti-GAPDH antibody; Abcam, Cambridge, UK) as reference proteins to control for adequate normalization.

Techniques: Control, Western Blot