Journal: Biochimica et biophysica acta

Article Title: 5'-AMP-activated protein kinase alpha regulates stress granule biogenesis.

doi: 10.1016/j.bbamcr.2015.03.015

Figure Lengend Snippet: Fig. 5. Compound C impairs SG formation. (A) HeLa cells were pre-incubated with the vehicle DMSO or CC for 1 h; DMSO or CC was also present throughout the subsequent treatment period. SG assembly was induced by arsenite; the vehicle water was added to controls. Alternatively, SGs were produced with DEM, whereas controls received ethanol. Both AMPK-α iso- forms (AMPK-α1/2) were detected in SGs demarcated with the marker HuR. (B) The same results as in part A were obtained with the SG marker G3BP1. Scale bar is 20 μm. (C) Western blots for control and DEM-treated samples show a reduction of α-subunit phosphorylation (α1, Thr183; α2, Thr172) after DEM incubation. CC further diminishes this modification. The loss of AMPK kinase activity was verified by probing for phosphorylated Acc1 (p-Acc1). Phosphorylation of eIF2α on Ser51 was not abolished by DEM or CC. DEM or CC had also no strong effects on the cellular concentrations of TIA-1/TIAR, G3BP1 or HuR.

Article Snippet: Primary antibodies were used at the following dilutions: AMPK-α1/2 (1:200), p-AMPK-α (1:2000; Cell Signaling #2535), p-Acc1 (1:1000; phosphoSer79; Cell Signaling #3661); Acc1 (1:500; Cell Signaling #3662), HuR (1:2000), G3BP1 (1:2000), TIA-1/TIAR (1:1000), AMPKα1 and AMPK-α2 (1:2000), and actin (1:100,000).

Techniques: Incubation, Produced, Marker, Western Blot, Control, Phospho-proteomics, Activity Assay

Journal: Biochimica et biophysica acta

Article Title: 5'-AMP-activated protein kinase alpha regulates stress granule biogenesis.

doi: 10.1016/j.bbamcr.2015.03.015

Figure Lengend Snippet: Fig. 11. AMPK activation precedes SG formation. HeLa cells were incubated with DEM for the time points indicated. (A) AMPK-α phosphorylation (p-AMPK-α1/2), total AMPK-α1/2, phosphorylated Acc1 (p-Acc1, Ser79) and total Acc1 were detected by Western blotting. Controls were incubated with the vehicle ethanol (EtOH). (B) The kinetics for SG formation was monitored by immunodetection of AMPK-α2 and G3BP1. SGs were formed after 60 min incubation with DEM. They contained the marker G3BP1 and AMPK-α2.

Article Snippet: Primary antibodies were used at the following dilutions: AMPK-α1/2 (1:200), p-AMPK-α (1:2000; Cell Signaling #2535), p-Acc1 (1:1000; phosphoSer79; Cell Signaling #3661); Acc1 (1:500; Cell Signaling #3662), HuR (1:2000), G3BP1 (1:2000), TIA-1/TIAR (1:1000), AMPKα1 and AMPK-α2 (1:2000), and actin (1:100,000).

Techniques: Activation Assay, Incubation, Phospho-proteomics, Western Blot, Immunodetection, Marker

Journal: The Journal of nutrition

Article Title: Cholic Acid Supplementation of a High-Fat Obesogenic Diet Suppresses Hepatic Triacylglycerol Accumulation in Mice via a Fibroblast Growth Factor 21-Dependent Mechanism.

doi: 10.1093/jn/nxy022

Figure Lengend Snippet: FIGURE 4 Abundance of P-ACC1 (Ser79) and total ACC1 in liver ex- tracts (A) and the abundance of P-HSL (Ser660 and Ser565) and total HSL in adipose extracts (B) of WT mice and FGF21KO mice fed an STD or an HFD for 23 wk or an HFD for 19 wk followed by 4 wk of HFD+CA. The ratio of P-ACC1 to total ACC1 and the ratio of P-HSL to total HSL in extracts prepared from WT mice fed the STD were set at 1 and the other values were adjusted proportionately. Values are means ± SEMs, n = 8. Labeled means without a common let- ter differ, P < 0.05. ACC1, acetyl-CoA carboxylase 1; CA, cholic acid; FGF21KO, fibroblast growth factor 21 knockout; HFD, high-fat obeso- genic diet; HSL, hormone-sensitive lipase; P-, phosphorylated; STD, standard diet; WT, wild-type.

Article Snippet: Antibodies against phosphorylated acetyl-CoA carboxylase 1 (ACC1) Ser79, phosphorylated hormone sensitive lipase (HSL) Ser565 and Ser660, total ACC1, and total HSL were obtained from Cell Signaling Technology.

Techniques: Labeling, Knock-Out

Journal: The Journal of nutrition

Article Title: Cholic Acid Supplementation of a High-Fat Obesogenic Diet Suppresses Hepatic Triacylglycerol Accumulation in Mice via a Fibroblast Growth Factor 21-Dependent Mechanism.

doi: 10.1093/jn/nxy022

Figure Lengend Snippet: FIGURE 4 Abundance of P-ACC1 (Ser79) and total ACC1 in liver ex- tracts (A) and the abundance of P-HSL (Ser660 and Ser565) and total HSL in adipose extracts (B) of WT mice and FGF21KO mice fed an STD or an HFD for 23 wk or an HFD for 19 wk followed by 4 wk of HFD+CA. The ratio of P-ACC1 to total ACC1 and the ratio of P-HSL to total HSL in extracts prepared from WT mice fed the STD were set at 1 and the other values were adjusted proportionately. Values are means ± SEMs, n = 8. Labeled means without a common let- ter differ, P < 0.05. ACC1, acetyl-CoA carboxylase 1; CA, cholic acid; FGF21KO, fibroblast growth factor 21 knockout; HFD, high-fat obeso- genic diet; HSL, hormone-sensitive lipase; P-, phosphorylated; STD, standard diet; WT, wild-type.

Article Snippet: Antibodies against phosphorylated acetyl-CoA carboxylase 1 (ACC1) Ser79, phosphorylated hormone sensitive lipase (HSL) Ser565 and Ser660, total ACC1, and total HSL were obtained from Cell Signaling Technology.

Techniques: Labeling, Knock-Out