Journal: Medicine & Science in Sports & Exercise

Article Title: Preexercise Aminoacidemia and Muscle Protein Synthesis after Resistance Exercise

doi: 10.1249/mss.0b013e31825d28fa

Figure Lengend Snippet: FIGURE 5—Muscle p70 S6 kinase thr421/ser424 (A), p70 S6 kinase thr389 (B), S6 ribosomal protein (C), and eEF2 kinase (D) phosphorylation at rest and after 1 and 5 h of recovery from a bout of resistance exercise in response to a placebo feeding (PLAC) or the intake of 30-g high-quality protein fed as a bolus (BOLUS) or in a series of small feedings (PULSE). Values are mean T SEM (n = 8) presented in arbitrary units relative to >-tubulin. *, different versus rest; ‡, different versus 5 h; #, different versus BOLUS— 1 h; § different versus PLAC—1 h (P G 0.05).

Article Snippet: Polyclonal anti–phospho-mammalian target of rapamycin (mTOR) ser2448 (#2971), -p70 S6K thr421/ser424 (#9204), -eEF2K ser366 (#3691); monoclonal anti–phospho-Akt ser473 (#9271), -tuberin (TSC2) thr1462 (#3617), -S6 ribosomal protein ser235/6 (#4856), and -p70 S6K thr389 (#9206) were from Cell Signaling Technology (Danvers, MA).

Techniques: Phospho-proteomics

Journal: PLoS ONE

Article Title: Inhibition of Elongation Factor-2 Kinase Augments the Antitumor Activity of Temozolomide against Glioma

doi: 10.1371/journal.pone.0081345

Figure Lengend Snippet: ( A ) Human glioma cell lines LN229 and U251 with or without silencing of eEF-2 kinase expression were plated in 96-well plates (5×10 3 /well) and treated with TMZ (100 μM) at 37°C for 24 h; ( B ) LN229 and U251 cells were plated in 96-well plates (5×10 3 /well) and treated with TMZ (100 μM) in the presence or absence of 0.5μM of NH125 at 37°C for 24 h. Cell viability was measured by CCK-8 assay. ( C ) LN229 and U251 cells were transfected with an eEF-2 kinase-targeted siRNA or a non-targeting control siRNA (NT control). At different time points, expression of eEF-2 kinase was analyzed by Western blot,. β-actin was used as a loading control. ( D ) LN229 and U251 cells were treated with NH125 (0.5μM) or vehicle for various periods of time. At the end of treatment, the level of phospho-eEF2 (Thr56) was examined by western blot. β-actin was used as a loading control. Each bar represents mean ± SD of triplicate determinations; results shown are the representative of three identical experiments; data are expressed as the percentage of the controls. ** p < 0.01.

Article Snippet: Temozolomide and dimethyl sulfoxide (DMSO) were purchased from Sigma (St Louis, MO); 1-Hexadecyl- 2-methyl-3-(phenylmethyl)-1H-imi-dazolium iodide (NH125) was obtained from Tocris Bioscience (St. Louis, MO); the antibodies to phospho-eEF2, eEF-2, casepase-3, PARP, and LC3B, were purchased from Cell Signaling Technology (Danvers, MA); rabbit polyclonal anti-eEF2 kinase antibody was obtained from Novus Biologicals (Littleton, CO); p62 was purchased from Enzo Life Sciences (Plymouth Meeting, PA); β-actin antibody was obtained from Santa Cruz Biotechnology Inc (Santa Cruz, CA); eEF-2 kinase-siRNA and control siRNA were synthesized by Shanghai Gene-Pharma Co. (Shanghai, China); the Cell Counting Kit-8 (CCK-8) was purchased from DojinDo Molecular Technologies, Inc. (Rockville, MA); the Annexin V-FITC apoptosis detection kit and Matrigel were purchased from BD Biosciences (San Diego, CA); the Pierce BCA Protein Assay Kit was obtained from Thermo Scientific Corp (Hudson, New Hampshire); oligofectamine reagent was purchased from Invitrogen Corp (Carlsbad, CA); other Western blot reagents were obtained from Bio-Rad Laboratories (Hercules, CA).

Techniques: Expressing, CCK-8 Assay, Transfection, Control, Western Blot