Journal: Physiological Reports

Article Title: Unilateral isometric contraction induces REDD1 and suppresses insulin‐stimulated mTORC1 and protein synthesis in non‐contracted muscle of male mice

doi: 10.14814/phy2.70574

Figure Lengend Snippet: Isometric contraction decreases insulin‐induced increase in mTORC1 signaling in non‐contracted muscle. (a) Representative immunoblots of phosphorylation of 4E‐BP1 and S6K1 (T389), respectively. (b, c) Changes in phosphorylation of 4E‐BP1 and S6K1, respectively, by the insulin and/or isometric contraction. In the PBS‐R and INS‐R groups, phosphorylation of 4E‐BP1 and S6K1 showed no significant differences between the left and right muscles of each mouse. Therefore, the average value of the left and right muscles is presented as a single bar with individual data points represented as dots in the figure (b, c). In the PBS‐IC and INS‐IC groups, each connected dot represents data obtained from the same animal. IC, isometrically contracted muscle; INS‐IC, mice treated with insulin and isometric contraction; INS‐R, mice treated with insulin (0.5 U/kg BW); N, non‐contracted muscle; PBS‐IC, mice treated with phosphate buffered saline and isometric contraction; PBS‐R, mice treated with phosphate‐buffered saline. The bars in the figure represent the mean ± SD, n = 5/group. Specific p values for significant differences are shown above the horizontal bars.

Article Snippet: Membranes were blocked for 1 h using Can Get Signal PVDF Blocking Reagent (Toyobo), then incubated overnight at 4°C with the following primary antibodies: phospho‐Thr389 S6K1 (#9234), total S6K1 (#2708), total 4E‐BP1 (#9644), phospho‐Thr308 Akt (#13038), phospho‐Ser473 Akt (#4060), and pan‐Akt (#4691) (Cell Signaling Technology); REDD1 (#10638‐1‐AP, Proteintech Group).

Techniques: Western Blot, Phospho-proteomics, Muscles, Saline

Journal: Physiological Reports

Article Title: Unilateral isometric contraction induces REDD1 and suppresses insulin‐stimulated mTORC1 and protein synthesis in non‐contracted muscle of male mice

doi: 10.14814/phy2.70574

Figure Lengend Snippet: Isometric contraction does not alter insulin‐induced increase in Akt phosphorylation in both contracted and non‐contracted muscle. (a) Representative immunoblots of phosphorylation of Akt at Thr308 and Ser473. (b, c) Changes in phosphorylation of Akt at Thr308 and Ser473, respectively, by the insulin and/or isometric contraction. In the PBS‐R and INS‐R groups, phosphorylation of 4E‐BP1 and S6K1 showed no significant differences between the left and right muscles of each mouse. Therefore, the average value of the left and right muscles is presented as a single bar with individual data points represented as dots in the figure (b, c). In the PBS‐IC and INS‐IC groups, each connected dot represents data obtained from the same animal. IC, isometrically contracted muscle; INS‐IC, mice treated with insulin and isometric contraction; INS‐R, mice treated with insulin (0.5 U/kg BW); N, non‐contracted muscle; PBS‐IC, mice treated with phosphate buffered saline and isometric contraction; PBS‐R, mice treated with phosphate‐buffered saline. The bars in the figure represent the mean ± SD, n = 5/group.

Article Snippet: Membranes were blocked for 1 h using Can Get Signal PVDF Blocking Reagent (Toyobo), then incubated overnight at 4°C with the following primary antibodies: phospho‐Thr389 S6K1 (#9234), total S6K1 (#2708), total 4E‐BP1 (#9644), phospho‐Thr308 Akt (#13038), phospho‐Ser473 Akt (#4060), and pan‐Akt (#4691) (Cell Signaling Technology); REDD1 (#10638‐1‐AP, Proteintech Group).

Techniques: Phospho-proteomics, Western Blot, Muscles, Saline

Journal: Drug Design, Development and Therapy

Article Title: Quantitative Proteomic Study Reveals Amygdalin Alleviates Liver Fibrosis Through Inhibiting mTOR/PDCD4/JNK Pathway in Hepatic Stellate Cells

doi: 10.2147/DDDT.S500439

Figure Lengend Snippet: Amygdalin upregulates PDCD4 via inhibiting activation mTOR/S6K1 signaling pathway. ( A ) LX-2 cells were stimulated with TGF-β1 for different time and MG132 for another 4 h. The protein level of PDCD4 was analyzed by Western blotting. ( B ) LX-2 cells were stimulated with TGF-β1 and the protein levels of total and phosphorylation levels of mTOR, S6K1 and PDCD4 were analyzed by Western blotting. ( C ) LX-2 cells were treated with amygdalin for 12 h or Rapamycin for 2 h and stimulated with TGF-β1 for another 24 h, and the protein levels of total and phosphorylated mTOR, S6K1, JNK, c-Jun were analyzed by Western blotting.

Article Snippet: The following primary antibodies: COL1A1 (CST, #72026), α-SMA (Abcam, ab5694), PDCD4 (CST, #9535), phospho-PDCD4 (Ser67) (Invitrogen, PA5-105015), mTOR (CST, #2972), phospho-mTOR (Ser2448) (CST, #5536), S6K1 (CST, #9202), phospho-S6K1 (Thr421/Ser424) (CST, #9204), JNK (Santa Cruz, sc-7345), phospho-JNK (Thr183/Tyr185) (Santa Cruz, sc-6254), c-Jun (CST, #9165), phospho-c-Jun (Ser73) (CST, #3270) and GAPDH (Sigma-Aldrich, G9545).

Techniques: Activation Assay, Western Blot, Phospho-proteomics