For this experiment, cells were pretreated with MG132 (20uM) for 1 hours. Surprisingly, I observed inhibitory effect of MG132 on protein expression. I have confirmed the MG132 effect by ubiquitin status. Autophagy inhibitor (CQ) was used to preclude the possibility of autophagy-mediated degradation. I did not observe any significant effect of CQ protein expression.
MG132 inhibits the proteolytic activity of the 26S proteasome complex that results in reduced degradation of ubiquitin-conjugated proteins. However, MG132 is reported to induce apoptosis in Hela cell with IC50 5 µM.You can follow the paper https://www.ncbi.nlm.nih.gov/pubmed/19513526
Hi Rani,
Can you please explain your experimental set-up in more details..?
Because, it is unclear in you current description.
And what do you mean by ---"Surprisingly, I observed inhibitory effect of MG132 on protein expression. I have confirmed the MG132 effect by ubiquitin status." and "I did not observe any significant effect of CQ protein expression".
Can you please elaborate?
Hi Rani
with the current description, it is hard to come to any sort of conclusion. It would be nice to have a detailed description of the experiment.
Abhishek
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