数据来源文献[1]. Cool B, et al. Identification and characterization of a small molecule AMPK activator that treats key components of type 2 diabetes and the metabolic syndrome. Cell Metab, 2006, 3(6), 403-416. [2]. Sanders MJ, et al. Defining the mechanism of activation of AMP-activate...
Cellular survival and function depend fundamentally on the liberation of free energy, achieved through the intermediary process of ATP hydrolysis, to drive anabolic processes. Cells have to contend with the challenge of fluctuating internal bioenergetic demands and environmental substrate availability, necessi...
21, activation of R299Q γ2 AMPK complexes by AMP was limited compared with WT (Fig.1c). In R299Q γ2 hearts, a corresponding increase in the phosphorylation of acetyl-CoA carboxylase at Ser79, a canonical AMPK target, was observed (Fig.1d;Supplementary Fig.1e), consistent with a bas...
The primary antibodies were mouse anti-α-tubulin monoclonal antibody (mAb) (Sigma-Aldrich), rat anti-α-tubulin mAb (Abcam), mouse anti-HA mAb (Covance), rat anti-HA mAb (Roche), and rat anti-GFP mAb (Nacalai Tesque) antibodies. Mouse anti-cingulin mAb and was gifted by K. Owaribe...
Rabbit monoclonal anti-p-AMPK Abcam Cat# ab32508; RRID: AB_722769 Rabbit monoclonal anti-AMPK Abcam Cat# ab32047; RRID: AB_722764 Rabbit polyclonal anti-FOXO3 Abcam Cat# ab23683; RRID: AB_732424 Rabbit polyclonal anti-SIRT3 Abcam Cat# ab189860 Rabbit monoclonal anti-BAX Abcam Cat# ab1827...
was used as a secondary antibody. The monoclonal anti-β-actin-peroxidase antibody (#A3854, Millipore Sigma, Co., St. Louis, MO, USA) was used as a loading control. Detection was performed using the ECL prime reagents (#RPN2232, Cytiva, Co., Tokyo, Japan) or Chemi-Lumi One Ultra (...
X-100 at 4 °C overnight as follows: Monoclonal rat anti-CD4 (diluted 1:50, BD Bioscience, San Jose, CA, USA) and CD8 (diluted 1:50, BD Bioscience, San Jose, CA, USA) antibodies or a polyclonal goat anti-CD20 antibody (diluted 1:50, SantaCruz Biotechnology, Dallas, TX, USA). ...
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Cells were blocked with CAS-block for 1 h. The cells were stained with rabbit monoclonal anti-TUFM antibody (abcam, USA) at 1:500 dilution for 1 h. Then the cells were washed with PBS and stained with antirabbit Alexa Flour 568 conjugated secondary antibody (Invitrogen, USA) for 1 h....
Monoclonal lines were expanded from single cells and their knockout status was then determined by western blot to use for subsequent experiments. Fluorescence imaging and image analysis For all live cell imaging experiments, cells were washed and incubated in Hanks balanced salt solution (HBSS, Gibco...