7974c  (Cell Signaling Technology Inc)

Journal: Molecules

Article Title: The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice

doi: 10.3390/molecules28041950

Figure Lengend Snippet: Effect of CP on myokine alterations. ( A – D ) The mRNA expression of myostatin ( A ), TGF-β ( B ), BDNF ( C ), and FNDC5 ( D ) in the muscles was assessed using real-time PCR. ( E , F ) Total Smad2/3, phospho-Smad2, total-AKT, phospho-AKT, and GAPDH levels in the muscles were assessed by immunoblotting. Total proteins were stained by Ponceau S. ImageJ was used to measure and quantify the band intensity. Arrow in ( E ) indicates p-Smad 2, and this band was used to quantify the p-Smad2 level. The p-Smad2 and p-AKT levels were normalized by Ponceau S staining. ( G ) Total and phospho-mTOR levels were analyzed by ELISA. The p-mTOR levels were normalized against the total mTOR levels. Data are presented as the means ± SD of values obtained from seven biological replicates. # p < 0.05, ## p < 0.01, DEX vs. control. # p < 0.05, ## p < 0.01, DEX + CP (0.25 and 0.5) vs. control. * p < 0.05, ** p < 0.01, DEX + CP (0.25 and 0.5) vs. DEX; CP, collagen peptides; DEX, dexamethasone.

Article Snippet: To quantify the activity of mTOR and p-mTOR, Pathscan sandwich ELISA kits (mTOR: CST#7974, p-mTOR: CST#7976) were used.

Techniques: Expressing, Muscles, Real-time Polymerase Chain Reaction, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecules

Article Title: The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice

doi: 10.3390/molecules28041950

Figure Lengend Snippet: Anti-inflammatory activity of CP in muscle tissue. ( A ) Evans blue staining of the calf muscle. The blue region in the representative images indicates damaged muscle fibers. The Evans blue-positive area was quantified and is presented as a relative percentage of the control below the representative photos. This experiment was performed with three mice per group. ( B , C ) The mRNA expression of TNF-α and IL-1β in the muscles was assessed using real-time PCR. Data are presented as the means ± SD of values obtained from seven biological replicates. ( D ) The IL-1β levels in serum were examined using ELISA. Data are presented as the means ± SD of values obtained from three biological replicates. # p < 0.05, ## p < 0.01, DEX vs. control. # p < 0.05, ## p < 0.01, DEX + CP (0.25 and 0.5) vs. control. * p < 0.05, ** p < 0.01, DEX + CP (0.25 and 0.5) vs. DEX; CP, collagen peptides; DEX, dexamethasone.

Article Snippet: To quantify the activity of mTOR and p-mTOR, Pathscan sandwich ELISA kits (mTOR: CST#7974, p-mTOR: CST#7976) were used.

Techniques: Activity Assay, Staining, Control, Expressing, Muscles, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Pharmaceuticals

Article Title: Novel mTORC1 Inhibitors Kill Glioblastoma Stem Cells

doi: 10.3390/ph13120419

Figure Lengend Snippet: Confirmation of mTORC1 inhibition by piperazine compounds. ( A ) Piperazine drugs dose-dependently inhibit mTORC1 downstream target S6Kinase in mouse FL83B cells. ( B ) Piperazine drugs inhibit GBM stem cells at a single concentration of 10 µM. The amount of phosphorylated S6Kinase relative to total S6Kinase was measured by ELISA. The bars are the fold change over the vehicle control. Error bars are 1 standard deviation. Data shown for meclizine, hydroxyzine, cinnarizine, and flunarizine as indicated. *, **, ***, **** correspond to p values less than 0.05, 0.001, 0.0001, and 0.00001 respectively. n = 3 for each group. p value calculated with Student’s t -test.

Article Snippet: The PathScan ® Phospho-S6Kinase (Thr389) Sandwich ELISA and PathScan ® Total S6K Sandwich ELISA Kits were purchased from Cell Signaling Technologies (Danvers, MA).

Techniques: Inhibition, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control, Standard Deviation