Journal: Frontiers in nutrition

Article Title: mTORC1 Signaling in Individual Human Muscle Fibers Following Resistance Exercise in Combination With Intake of Essential Amino Acids.

doi: 10.3389/fnut.2019.00096

Figure Lengend Snippet: FIGURE 2 | Protein levels in individual type I and type II fibers before exercise from the five subjects. (A) mTOR, (B) S6K1, and (C) eEF2. Horizontal lines illustrate the average values.

Article Snippet: The primary antibodies used were Mouse Anti-Slow Skeletal Myosin Heavy Chain (Abcam #ab11083, diluted 1:10,000), Rabbit Anti-Fast Myosin Skeletal Heavy Chain (Abcam #ab91506, diluted 1:10,000), total mTOR (Cell Signaling #2983S, diluted 1:1,000), phosphorylated mTOR Ser2448 (Cell Signaling #5536S, diluted 1:1,000), total S6K1 (Cell Signaling #2708S, diluted 1:1,000), phosphorylated S6K1 Thr389 (Cell Signaling #9234S, diluted 1:1,000), total eEF2 (Cell Signaling #2332S, diluted 1:1,000), and phosphorylated eEF2 Thr56 (Cell Signaling #2331S, diluted 1:1,000).

Techniques:

Journal: Frontiers in nutrition

Article Title: mTORC1 Signaling in Individual Human Muscle Fibers Following Resistance Exercise in Combination With Intake of Essential Amino Acids.

doi: 10.3389/fnut.2019.00096

Figure Lengend Snippet: FIGURE 1 | Identification of type I and type II fibers following incubation with antibodies targeting MHCI and MHCII. Lower bands represent protein levels of mTOR, S6K1, and eEF2 in individual fibers. Proteins were separated on 4–20% acrylamide gels and transferred to PVDF membranes.

Article Snippet: The primary antibodies used were Mouse Anti-Slow Skeletal Myosin Heavy Chain (Abcam #ab11083, diluted 1:10,000), Rabbit Anti-Fast Myosin Skeletal Heavy Chain (Abcam #ab91506, diluted 1:10,000), total mTOR (Cell Signaling #2983S, diluted 1:1,000), phosphorylated mTOR Ser2448 (Cell Signaling #5536S, diluted 1:1,000), total S6K1 (Cell Signaling #2708S, diluted 1:1,000), phosphorylated S6K1 Thr389 (Cell Signaling #9234S, diluted 1:1,000), total eEF2 (Cell Signaling #2332S, diluted 1:1,000), and phosphorylated eEF2 Thr56 (Cell Signaling #2331S, diluted 1:1,000).

Techniques: Incubation

Journal: Frontiers in nutrition

Article Title: mTORC1 Signaling in Individual Human Muscle Fibers Following Resistance Exercise in Combination With Intake of Essential Amino Acids.

doi: 10.3389/fnut.2019.00096

Figure Lengend Snippet: FIGURE 3 | (A) Phosphorylation of mTOR at Ser2448, (B) total protein level of mTOR, (C) phosphorylation of mTOR/total protein, (D) phosphorylation of S6K1 at Thr389, (E) total protein level of S6K1, (F) phosphorylation of S6K1/total protein, (G) phosphorylation of eEF2 at Thr56, (H) total protein level of eEF2, and (I) phosphorylation of eEF2/total protein. Pre indicates before exercise and 90 indicates 90 min after exercise. Figures within bars indicate the number of fibers analyzed. The values presented (arbitrary units/100) are means ± SE for the five subjects. *P < 0.05 vs. Pre, #P < 0.05 vs. placebo, and $P < 0.05 vs. type I fibers.

Article Snippet: The primary antibodies used were Mouse Anti-Slow Skeletal Myosin Heavy Chain (Abcam #ab11083, diluted 1:10,000), Rabbit Anti-Fast Myosin Skeletal Heavy Chain (Abcam #ab91506, diluted 1:10,000), total mTOR (Cell Signaling #2983S, diluted 1:1,000), phosphorylated mTOR Ser2448 (Cell Signaling #5536S, diluted 1:1,000), total S6K1 (Cell Signaling #2708S, diluted 1:1,000), phosphorylated S6K1 Thr389 (Cell Signaling #9234S, diluted 1:1,000), total eEF2 (Cell Signaling #2332S, diluted 1:1,000), and phosphorylated eEF2 Thr56 (Cell Signaling #2331S, diluted 1:1,000).

Techniques: Phospho-proteomics

Journal: Physiological Reports

Article Title: Moderate alcohol consumption does not impair overload-induced muscle hypertrophy and protein synthesis

doi: 10.14814/phy2.12333

Figure Lengend Snippet: Muscle overload-induced increases in S6K1 substrates rpS6 and eEF2 are not prevented by alcohol consumption. The relative amount of phosphorylated and total rpS6 (A, B) and eEF2 (C, D) were measured in the plantaris muscle after 14 days of overload. Representative images of each marker correspond to the sample order depicted in the graph. * P < 0.05, indicates difference from Sham condition within that treatment. Values are expressed as means ± SE.

Article Snippet: Antibodies included (Cell Signaling, Beverly, MA, unless otherwise noted): S6K1, S6K1 (Thr 389 ), rpS6, rpS6 (Ser 240/244 ), 4E-BP1 (Bethyl Laboratories, Montgomery, TX), 4E-BP1 (Thr 37/46 ), eEF2, eEF2 (Thr 56 ), ERK1/2 (Thr 202 /Tyr 204 ), p42/44 MAPK, mTOR, mTOR (Ser 2448 ), REDD1 (ProteinTech, Chicago, IL), Akt, Akt (Thr 308 ), PRAS40, PRAS40 (Thr 246 ), ULK1, ULK1 (Ser 757 ), p62, LC3A/B, and GAPDH.

Techniques: Marker