Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Neuroprotective Impact of Linagliptin against Cadmium-Induced Cognitive Impairment and Neuropathological Aberrations: Targeting SIRT1/Nrf2 Axis, Apoptosis, and Autophagy.

doi: 10.3390/ph16081065

Figure Lengend Snippet: Figure 8. Linagliptin stimulates hippocampal AMPK/mTOR pathway in cadmium-intoxicated rats. Herein, stimulation of AMPK/mTOR pathway by linagliptin in cadmium-intoxicated animals was reflected by the elevation in p-AMPK/AMPK ratio (B), and reduction in p-mTOR/mTOR ratio (D). Of note, the individual O.D. of p-AMPK(Ser487) and total AMPK are shown in (A), whereas the individual O.D. of p-mTOR(Ser2448) and total mTOR are shown in (C). For each experimental group, the mean optical density of either p-AMPK or p-mTOR was divided by the corresponding mean optical density of the total AMPK or total mTOR, respectively. This was followed by setting the mean control value to 1. N = 6 in each group (mean ± standard error of the mean). A p-value of less than 0.05 was significant. * p < 0.05, ** p < 0.01, or **** p < 0.0001, compared to control; # p < 0.05, or ## p < 0.01, compared to cadmium (Tukey’s test for multi-comparisons and one-way ANOVA). AMPK, 5′adenosine-monophosphate-activated protein kinase/ mammalian target of rapamycin; Cd, cadmium chloride; LIN, linagliptin; mTOR, mammalian target of rapamycin.

Article Snippet: Likewise, ELISA kits from Cell Signaling were used to quantify phosphorylated and total forms of mTOR (Cat. # 7976C for p-mTOR[Ser2448] and Cat. # 7974C for pan mTOR, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Control

Journal:

Article Title: Aerobic Exercise Overcomes the Age-Related Insulin Resistance of Muscle Protein Metabolism by Improving Endothelial Function and Akt/Mammalian Target of Rapamycin Signaling

doi: 10.2337/db06-1566

Figure Lengend Snippet: Phosphorylation of Akt/PKB, mTOR, 4E-BP1, and S6K1 in two groups of older subjects at baseline and during insulin infusion performed ~20 h following rest (Control: n = 5 for Akt/PKB and mTOR; n = 4 for 4E-BP1 and S6K1) or aerobic exercise (Exercise: n = 6). We used the following phospho and total primary antibodies (Cell Signaling, Beverly, MA): phospho-mTOR (Ser2448, cat. no. 2971, lot no. 9; 1:1,000), phospho-p70 S6K1 (Thr389, cat. no. 9234, lot no. 2; 1:500), phospho-Akt (Ser473, cat. no. 4058, lot no. 6; 1:500), and phospho-4EBP1 (Thr37/46, cat. no. 2971; 1:1,000). Anti-rabbit IgG horseradish peroxidase–conjugated secondary antibody was purchased from Amersham Bioscience (1:2,000). Values are means ± SE. *P < 0.05 vs. basal, #P < 0.05 vs. control. □, basal; ■, insulin. Bas, basal; Ins, insulin; MW, molecular weight.

Article Snippet: We used the following phospho and total primary antibodies (Cell Signaling, Beverly, MA): phospho-mTOR (Ser 2448 , cat. no. 2971, lot no. 9; 1:1,000), phospho-p70 S6K1 (Thr 389 , cat. no. 9234, lot no. 2; 1:500), phospho-Akt (Ser 473 , cat. no. 4058, lot no. 6; 1:500), and phospho-4EBP1 (Thr 37/46 , cat. no. 2971; 1:1,000).

Techniques: Phospho-proteomics, Control, Molecular Weight

Journal: Oncology Letters

Article Title: Luteolin reduces migration of human glioblastoma cell lines via inhibition of the p-IGF-1R/PI3K/AKT/mTOR signaling pathway

doi: 10.3892/ol.2017.6643

Figure Lengend Snippet: The p-IGF-1R/phosphoinositide 3 kinase/AKT/mTOR signaling pathway was involved in the anti-migration progress of luteolin. (A) Cells were exposed to various concentrations of luteolin for 24 h and the expression of p-IGF-1R, p-AKT, AKT, p-mTOR and mTOR was detected by western blot. (B) U251MG and U87MG cells were serum-starved overnight and then treated for 24 h with 20 µM luteolin prior to 1 h stimulation with 100 ng/ml IGF-1. The expression of p-IGF-1R, p-AKT, AKT, p-mTOR, mTOR, MMP-2, MMP-9, TIMP-1, TIMP-2, E-cadherin, Vimentin and β-actin was determined. p-, phosphorylated; IGF-1R, insulin-like growth factor-1 receptor; AKT, protein kinase B; mTOR, mammalian target of rapamycin; IGF-1, insulin-like growth factor-1; MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinase.

Article Snippet: Anti-E-cadherin, anti-N-cadherin, anti-vimentin, anti-β-catenin, anti-vimentin (EMT kit; cat. no. cst-9782; 1:1,000), anti-p-protein kinase B (AKT) (cst-4060; 1:1,000), anti-AKT (cat. no. cst-9272; 1:1,000), anti-p-mammalian target of rapamycin (mTOR) (cat. no. cst-2971; 1:1,000), anti-mTOR (cat. no. cst-2983; 1:1,000), anti-β-actin (cat. no. cst-4970; 1:1,000), and horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG; heavy and light chain; cat. no. cst-7074; 1:5,000) secondary antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Migration, Expressing, Western Blot