I got the increased pattern of IRE1 alpha in western blot, whereas the expression of XBP-1 is decreased when compared with p-IRE1. I wonder, whether IRE1alpha is XBP-1 independent. Moreover, there was no any differences in mRNA where i checked for the spliced form of XBP-1.
Hi Kashi,
First, XBP1 mRNA expression is affected by N-ATF6, but not IRE1a or p-IRE1a. If your experiments of ER stress model will not affect ATF6 activity, XBP1 expression will not change. In my experiments, Tunicamycin (TM) would trigger all UPR activity (IRE1a, ATF6, and PERK). You may use this drug to test ER stress in your experiments.
XBP1s from XBP1 is regulated by p-IRE1a. If your data present highly p-IRE1a, your XBP1s (mRNA or protein) should be increased.
BIP is also regulated by ATF6. You also can test BIP mRNA and protein to verify ATF6 activity.
Luke TH Tsai
Thank you Dr. Tsung for your quick reply. ATF6a and IRE1a have similar expression with increasing pattern. I am doing invivo experiments so did not use Tunicamycin. Almost all the ER stress related proteins including BiP has been increased except XBP-1.
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