Discovery of autophagy modulators through the construction of a high-content screening platform via monitoring of lipid droplets

2013 ◽  
Vol 4 (8) ◽  
pp. 3282 ◽  
Author(s):  
Sanghee Lee ◽  
Eunha Kim ◽  
Seung Bum Park
Author(s):  
Amit Paul ◽  
David Franz ◽  
Sumaira Yahya ◽  
Shan Sun ◽  
Michael Cho

Recent evidence suggests that stem cell differentiation can be regulated by modulation of the cell’s biomechanics. The cytoskeletal structures and arrangements in stem cells undergoing differentiation are dramatically altered, and these alterations vary by lineage. The complexity of events associated with the transformation of these precursor cells leaves many questions unanswered about morphological, structural, proteomic, and functional changes in differentiating stem cells. A thorough understanding of stem cell behavior, both experimentally and computationally, would allow for the development of more effective approaches to the expansion of stem cells in vitro and for the regulation of their commitment to a specific phenotype.


2008 ◽  
Vol 13 (9) ◽  
pp. 837-846 ◽  
Author(s):  
Michael Wolff ◽  
Stefan G. Kauschke ◽  
Susanne Schmidt ◽  
Ralf Heilker

In the liver, glucokinase (GK) regulatory protein (GKRP) negatively modulates the metabolic enzyme GK by locking it in an inactive state in the nucleus. Here, the authors established a high content screening assay in the 384-well microplate format to measure the nucleus-to-cytoplasm translocation of GK by reagents that destabilize the interaction between GK and GKRP. As a cellular model system, primary rat hepatocytes endogenously expressing both GK and GKRP at physiological levels were used. The GK translocation assay was robust, displayed limited day-to-day variability, and delivered good Z′ statistics. The increase of the glucose concentration in the extracellular medium from a low glucose situation (2.8 mM) to beyond its physiological set point value of 5 mM was found to drive GK from the nucleus into the cytoplasm. Likewise, both fructose (converted intracellularly into fructose-1-phosphate) and a known allosteric GK activator were found to induce the export of GK from the nucleus and to synergistically enhance the effects of medium or high glucose concentrations with respect to GK translocation. Transfer of the high content screening format to a semiautomated medium throughput screening platform enabled the profiling of large compound numbers with respect to allosteric activation of GK. ( Journal of Biomolecular Screening 2008:837-846)


2012 ◽  
Vol 17 (5) ◽  
pp. 359-369 ◽  
Author(s):  
Constantin Radu ◽  
Hosna Sana Adrar ◽  
Ab Alamir ◽  
Ian Hatherley ◽  
Trung Trinh ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document