The cell cytosol is characterized by a high degree of macromolecular crowding, which profoundly affects protein–protein interactions15. To investigate how the available in vitro data apply to the situation in the intact cell, here we explored the chaperonin mechanism within its natural cellular contex...
Persistent endothelial injury promotes maladaptive responses by favoring the release of factors leading to perturbation in vascular homeostasis and tissue architecture. Caspase-3 dependent death of microvascular endothelial cells leads to the release of
islet β-cell control of Proinsulin translation remains incompletely understood. Here, we identify OSGEP, an enzyme responsible for t6A37modification of tRNANNUthat tunes glucose metabolism in β-cells. Global Osgep deletion causes glucose intolerance, while β-cell-specific deletion induces hyperglycemia ...
The TAR DNA Binding Protein 43 (TDP-43) has been implicated in the pathogenesis of human neurodegenerative diseases and exhibits hallmark neuropathology in amyotrophic lateral sclerosis (ALS). Here, we explore its tractability as a plasma biomarker of di
(SDS-PAGE) and protein staining (observe below). Gel electrophoresis and immunoblotting. Samples were treated with 10 sample buffer (125 mM Tris-HCl, 10% SDS, 20% glycerol, 5% 2-mercaptoethanol, 0.01% bromophenol blue) for 2 min at 100C before loading onto the polyacrylamide gels. ...
Nuclear transport factor 2 and small GTPase Ran participate in the nucleo-cytoplasm transport of macromolecules, but their function in the 20-hydroxyecdysone (20E) signal transduction pathway are not well known. A 703 bp encoding Ntf2 and a 1233 bp encod
SDS-PAGE of isolated and solubilized egg coats with 20% beta-mercaptoethanol or 25 mM dithiothreitol (DTT) showed bands between 58 and 105 kDa and between 44 and 103 kDa, respectively. During normal fertilization, the sperm undergoes a drastic change in shape after first contact with the egg....
Cells were washed with phosphate-buffered saline (PBS) and homogenized in cell lysis buffer (150 mM NaCl, 1% NP40, 10% DOC, 10% SDS, 50 mM Tris, pH 7.4). Determination of protein concentration was performed using the BCA protein assay kit (Pierce). Samples were mixed with standard Laemm...
Cell lysates were mixed with Laemmli buffer (Bio-Rad, cat# 1610747) containing β-mercaptoethanol, and boiled at 95 °C for 5 minutes and subjected to SDS–PAGE. Proteins were transferred onto PVDF membranes and incubated overnight with primary antibodies against SMS (1:500, clone 1G6, ...
The nature and assembly of the chlamydial division septum is poorly defined due to the paucity of a detectable peptidoglycan (PG)-based cell wall, the inhibition of constriction by penicillin and the presence of coding sequences for cell wall precursor a