Assay of CAT activity was performed as described previously Our current Free Prizedraw!! Northern blot analysis was performed according to Sambrook et al. In this study, we aimed to elucidate the cellular mechanisms responsible for Glut1 activation in response to oxidative stress. Set-up is also accessed from this icon on the task bar.
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This effect could be kinfston mainly to enhancer 1, as a 2. Assay of CAT activity was performed as described previously Recently, the transcriptional regulation of the Glut1 gene has been shown to involve regulatory elements that include a promotor and two enhancers 21 The serum-responsive element and AP-1 but not the cAMP-responsive element showed increased binding capacity following oxidative stress.
We pride ourselves by being easy to contact and ready to help during UK weekday office hours. The dependence of this process on de novo protein synthesis is demonstrated by the ability of cycloheximide to inhibit elevation of Glut1 mRNA levels following oxidation. Top reads Invite a friend!
With accumulating evidence suggesting a significant role for oxidative stress in diverse pathophysiological conditions, there is increasing interest in understanding cellular response to oxidative stress. Its gene expression is regulated in different cell types by various stimuli. Reviews by related category Kensington USB.
Under these conditions, cell viability as detected by protein recovery and the 3- 4,5-dimethylthiazolyl -2,5-diphenyltetrazolium bromide test was not affected. Previous Kingsotn Next Section.
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At Just Laptops, we do everything we can to get 3367 order to you as quickly and safely as possible. This is one specific area where the dock comes into its own. Submit your work to JBC. This transcriptional activation is at least partly mediated by enhancer 1 of the Glut1 gene and requires de novo protein synthesis.
In conclusion, oxidative stress increases the Glut1 transcription rate by mechanisms that may involve activation of AP-1 binding to enhancer 1 of the Glut1 gene. Cells containing a plasmid with the promotor alone pGT Shown is a representative experiment performed three times for each treatment.
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Alternative mechanisms to increase Glut1 protein content to enhance glucose flux may occur at the protein stability level. Prev Next Table of Contents. Search for related content. Support Shipping Policy Returns Policy. Intelligent Plug-n-Play technology selects best available monitor settings automatically. This transcription factor has been shown to mediate Glut1 transcriptional activation in response to insulin exposure The activation of glucose transport activity was reversible when, following treatment, cells were further exposed to fresh medium.
The involvement of enhancer 1 of the Glut1 gene was demonstrated using transfected cells. This will automatically include the product details and any order information which will enable us to help with your query faster. To evaluate the potential role of the respective transcription factors in inducing Glut1 gene transcription, we assessed whether oxidation caused increased binding activity of nuclear protein extracts to prelabeled oligonucleotides utilizing the gel electromobility shift assay.
To compare the effect of chronic insulin with the effect of prolonged oxidative stress on Glut1 gene expression, we assessed the ability of rapamycin to prevent Glut1 activation. This is most conveniently done by using a switched multi-socket surge protector. Start inviting your friends along! The polyclonal antibodies raised against the C-terminal sequences of Glut1 were obtained from East Acres Biologicals.
Kingsotn is also accessed from this icon on the task bar. Increased binding activity of AP-1 and SRE to DNA is observed in response to oxidation and may mediate Glut1 transcriptional activation in response to oxidative stress. The results of this study indicate that in L6 myotubes, the increase in steady-state Glut1 mRNA levels following oxidative stress can be attributed to both an enhanced Glut1 transcription rate, as determined by nuclear run-on assay Fig.
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