Journal: Poultry science

Article Title: The influence of dietary leucine above recommendations and fixed ratios to isoleucine and valine on muscle protein synthesis and degradation pathways in broilers.

doi: 10.3382/ps/pez396

Figure Lengend Snippet: Figure 7. Relative phosphorylation levels (fold of L0) of mTOR (A), S6K1 (B), and rpS6 (C) in the breast muscle of broilers fed either control diets (L0) or diets supplemented with moderate (L1) or high (L2) concentrations of leucine at fixed ratios of leucine: isoleucine and leucine: valine at days 10, 21, and 35 (mean ± SE). Relative phospho- rylation levels were calculated as the amount of protein detected by the phosphorylated antibody divided by that detected by the antibody for detection of total protein level of the respective protein. mTOR, mammalian target of rapamycin; S6K1, ribosomal protein S6 kinase beta-1, rpS6, ribosomal protein S6. The data set was analyzed by 1- way ANOVA with leucine concentration as fixed factor and the Tukey test was used as post hoc test. The experimental unit was the individ- ual animal (n = 4–5). There were no significant (P < 0.05) differences between experimental groups. Representative blots for one animal of each group at day 35 are shown.

Article Snippet: The membranes were incubated with primary antibodies against total and phosphorylated Akt (Cat. 4685 and 4051, CST, Danvers, MA), FOXO1 (Cat. 9454 and 9461, CST), eIF2α (Cat. 9722 and 9721, CST), mTOR (Cat. 2972 and 2971, CST), S6 ribosomal protein (rpS6) (Cat. 2217 and 2211, CST), and S6K1 (Cat. 9202 and 9234, CST), at a dilution of 1:1000 in 5% skimmed milk powder in 1 x TBST for phosphorylated Akt and in 5% BSA in 1 x TBST for the other antibodies at 4◦C overnight.

Techniques: Phospho-proteomics, Control, Concentration Assay

Journal: Scientific Reports

Article Title: Regulation of filial imprinting and structural plasticity by mTORC1 in newborn chickens

doi: 10.1038/s41598-018-26479-1

Figure Lengend Snippet: Experience-dependent activation of mTORC1. ( a ) Tissue samples from IMM (green) and MNM (blue) were collected for western blotting, 15 minutes after training. ( b ) Phosphorylation of the S6 ribosomal protein in MNM (left) and IMM (right) was increased in trained (N = 7) chickens compared to controls (N = 6). Representative western blots are shown below each panel. Box plots show mean and interquartile. Error bars show 10–90 percentile, *indicates p < 0.05 from two-sample t-test. The number of animals used in each group is listed inside parenthesis. (Illustrations by Michael Beckert).

Article Snippet: We used antibodies against the phosphorylated ribosomal protein p-s6 (Cell Signaling Cat #2215) and total s6 protein (Cell Signaling Cat #2217), following standard protocols described before .

Techniques: Activation Assay, Western Blot, Phospho-proteomics