Journal: PLoS ONE

Article Title: The Mechanism of Action of the Histone Deacetylase Inhibitor Vorinostat Involves Interaction with the Insulin-Like Growth Factor Signaling Pathway

doi: 10.1371/journal.pone.0024468

Figure Lengend Snippet: Ishikawa and USPC-2 cells were treated with vorinostat and/or IGF-I, and cell lysates (100 µg) were prepared after 24 h. Western blot analysis was performed with antibodies against pmTor, T-mTor, p-Ampk, T-Ampk, p21, cyclin D1, PI3K and actin. The figure shows the results of a typical experiment, repeated three times with similar results.

Article Snippet: Antibodies against phospho-IGF-IR (3024), IGF-IR β-subunit (3027), insulin receptor [(IR); 3025], phospho-AKT (9271), AKT (9272), phospho-ERK1/2 (9106), poly ADP ribose polymerase [(PARP); 9542], caspase 3 (9661), phospho-Ampk (2531), Ampk (2532), phospho-mTor (5536), mTor (2983), and PI3K p85 (4292) were obtained from Cell Signaling Technology.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: The Metabolic Consequences of Hepatic AMP-Kinase Phosphorylation in Rainbow Trout

doi: 10.1371/journal.pone.0020228

Figure Lengend Snippet: Fish were fasted (48 h) prior to injection and livers were sampled 2 h after injection. Gels were loaded with 20 µg total protein per lane. Protein (total AMPK) and phosphorylation levels (AMPK Thr 172 ) were normalized to tissue β-tubulin levels. Results are expressed as means ± s.e.m. ( n = 6) and were analyzed by one-way ANOVA followed by Student-Newman-Keuls comparison test. *Significant difference ( α <0.05).

Article Snippet: Protein extraction (20 µg) and Western blotting were undertaken as in using anti-phospho-AMPKα Thr 172 and anti-AMPK antibodies (Cell Signaling Technology, France) that recognize both α-1 and α-2 isoforms of the enzyme.

Techniques: Injection

Journal: PLoS ONE

Article Title: The Metabolic Consequences of Hepatic AMP-Kinase Phosphorylation in Rainbow Trout

doi: 10.1371/journal.pone.0020228

Figure Lengend Snippet: Gels were loaded with 20 µg total protein per lane. Protein (total AMPK) and phosphorylation levels (AMPK Thr 172 ) were normalized to tissue β-tubulin levels. mRNA levels were estimated using real-time RT-PCR. Expression levels were normalized to β-actin-expressed transcripts which did not change under the experimental conditions and are presented as fold-change against the saline solution-treated group set to 1. Results are expressed as means ± s.e.m. ( n = 6) and were analyzed by one-way ANOVA followed by Student-Newman-Keuls comparison test. *Significant difference ( α <0.05).

Article Snippet: Protein extraction (20 µg) and Western blotting were undertaken as in using anti-phospho-AMPKα Thr 172 and anti-AMPK antibodies (Cell Signaling Technology, France) that recognize both α-1 and α-2 isoforms of the enzyme.

Techniques: Quantitative RT-PCR, Expressing