Journal:

Article Title: THE ALTERNATIVE STIMULATORY G PROTEIN ?-SUBUNIT XL?s IS A CRITICAL REGULATOR OF ENERGY AND GLUCOSE METABOLISM AND SYMPATHETIC NERVE ACTIVITY IN ADULT MICE *

doi: 10.1074/jbc.M511752200

Figure Lengend Snippet: Blood glucose in Gnasxlm+/p− mice (□, dashed lines) and wild-type littermates (▪, solid lines) measured at indicated time points before and after A, glucose or B, insulin administration (n = 4-5 per group; results for insulin tolerance test expressed as % of baseline). Genotype had a significant effect by two-factor ANOVA for both studies. C, Basal (pre-clamp) and clamp endogenous glucose production rate, glucose infusion rate, and rates of whole body glucose uptake, glycolysis, and glycogen synthesis in wild-type (closed bars) and Gnasxlm+/p− mice (open bars) during hyperinsulinemic-euglycemic clamp studies (n = 7/group). Basal and clamp glucose (top panel) and insulin levels (bottom panel) are shown to the right. D, Glucose uptake rates in gastrocnemius muscle, WAT, and BAT, respectively, during the clamp study. E, Glycogen content of hindlimb muscles obtained from non-fasting wild-type (open bar) and mutant (closed bar) mice (n = 5–7/group). F, Results of immunoblotting using anti-Akt1 phospho-Ser473 (top of each panel) followed by anti-Akt Ab (bottom of each panel) in BAT, WAT, and muscle tissue extracts derived from wild-type (WT) and Gnasxlm+/p− (XL) mice which were sacrificed at indicated times after insulin administration. G, Serum C-peptide/insulin ratios from non-fasted 8 mo old male mice (n = 5-6/group). *p < 0.05 or **p < 0.01 vs. wild-type.

Article Snippet: Immunoprecipitations were performed on tissue extracts (1 mg protein) using an anti-Akt1 Ab (Upstate Biotechnology, Lake Placid, NY) as per manufacturer’s instructions, followed by immunoblot analysis using an anti-Akt1 phospho-Ser 473 Ab (Upstate).

Techniques: Muscles, Mutagenesis, Western Blot, Derivative Assay

Journal:

Article Title: Multiple signalling pathways involved in β 2 -adrenoceptor-mediated glucose uptake in rat skeletal muscle cells

doi: 10.1038/sj.bjp.0706626

Figure Lengend Snippet: In (a) [3H]-2-DG uptake (GU) in response to insulin (1 μM) or zinterol (1 μM) was significantly inhibited by the presence of PI3K inhibitor LY294002 (1 μM) (Student's t-test, P=0.01 and P<0.001, n=4–5), whereas the response to 8Br-cAMP (1 mM) was not affected by LY294002 (P=0.9, n=5). In (b) Western blot analysis showed that phosphorylation of Akt at Ser473 occurred in response to insulin (10 μM; 5 min stimulation) but not zinterol (100 nM; 3 h stimulation). Total Akt was detected in all samples. The labels are: basal (1), zinterol (2) and insulin (3). Blots from three separate experiments are shown.

Article Snippet: Samples were harvested in prewarmed (65°C) sodium dodecyl sulfate sample buffer containing 50 m M dithiothreitol, sonicated, boiled for 3 min, and separated on 5% acrylamide gel (or 10% for Akt) for 2 h at 100 V. The proteins were transferred on to Hybond-C extra nitrocellulose (Amersham Biosciences UK Ltd, U.K.) or PVDF membrane (Millipore, Australia), and probed with primary antibodies: phosphor-acetyl-CoA carboxylase (Ser79), (dissolved 1 : 1000 in 5% BSA), phospho-Akt (Ser473) and total Akt (1 : 2000 in 5% BSA) (Cell Signalling Technology, Inc., Danvers, MA, U.S.A.).

Techniques: Western Blot

Journal: British Journal of Cancer

Article Title: Phosphorylation of the androgen receptor is associated with reduced survival in hormone-refractory prostate cancer patients

doi: 10.1038/sj.bjc.6604152

Figure Lengend Snippet: Histoscore variation and comparison of staining intensity for hormone-sensitive and hormone-refractory tumours

Article Snippet: Phosphatidylinositol 3-OH kinase (Cell Signalling Technology, Beverly, MA, USA), Akt1–3 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), pAkt 473 (Cell Signalling Technology), mTOR (Santa Cruz Biotechnology Inc.), pmTOR 2448 (Cell Signalling Technology) and pAR 210 (Imgenex, San Diego, CA, USA) antibodies were used at the following concentrations (1, 1, 2, 2, 4, 5, 2 and 50 μ g ml −1 ).

Techniques: Staining

Journal: British Journal of Cancer

Article Title: Phosphorylation of the androgen receptor is associated with reduced survival in hormone-refractory prostate cancer patients

doi: 10.1038/sj.bjc.6604152

Figure Lengend Snippet: ( A ) Kaplan–Meier plot demonstrates that those patients whose tumours exhibit a rise in PI3K expression (broken line) relapse quicker than those patients whose tumours exhibit no change or a fall in PI3K expression (solid line). ( B ) Kaplan–Meier plot demonstrates that those patients whose tumours exhibit a rise in pAR 210 expression (broken line) have shorter time to disease-specific death from time of biochemical relapse than those patients whose tumours exhibit no change or a fall in pAR 210 expression (solid line). ( C ) Kaplan–Meier plot demonstrates that those patients whose tumours exhibit a rise in pAkt 473 cytoplasmic expression (broken line) have shorter time to disease-specific death than those patients whose tumours exhibit no change or a fall in pAkt 473 expression (solid line). ( D ) Kaplan–Meier plot demonstrates that those patients whose tumours exhibit a rise in pAR 210 expression (broken line) have shorter time to disease-specific death than those patients whose tumours exhibit no change or a fall in pAR 210 expression (solid line).

Article Snippet: Phosphatidylinositol 3-OH kinase (Cell Signalling Technology, Beverly, MA, USA), Akt1–3 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), pAkt 473 (Cell Signalling Technology), mTOR (Santa Cruz Biotechnology Inc.), pmTOR 2448 (Cell Signalling Technology) and pAR 210 (Imgenex, San Diego, CA, USA) antibodies were used at the following concentrations (1, 1, 2, 2, 4, 5, 2 and 50 μ g ml −1 ).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Phosphorylation of the androgen receptor is associated with reduced survival in hormone-refractory prostate cancer patients

doi: 10.1038/sj.bjc.6604152

Figure Lengend Snippet: Scatter plots of pAkt 473 histoscore compared to pAR 210 histoscore, ( A ) is in the hormone-sensitive tissue and no significant correlation was observed ( P =0.061, correlation coefficient 0.251); however, ( B ) is in the hormone-refractory tissue, where a significant correlation was observed ( P <0.001 and correlation coefficient 0.711).

Article Snippet: Phosphatidylinositol 3-OH kinase (Cell Signalling Technology, Beverly, MA, USA), Akt1–3 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), pAkt 473 (Cell Signalling Technology), mTOR (Santa Cruz Biotechnology Inc.), pmTOR 2448 (Cell Signalling Technology) and pAR 210 (Imgenex, San Diego, CA, USA) antibodies were used at the following concentrations (1, 1, 2, 2, 4, 5, 2 and 50 μ g ml −1 ).

Techniques: