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. Author manuscript; available in PMC: 2010 Aug 7.
Published in final edited form as: Cell. 2009 Aug 7;138(3):562–575. doi: 10.1016/j.cell.2009.07.017

Figure 1. Conditional Tools to Forcibly Trigger IRE1α Catalytic Activities.

Figure 1

(A) IRE1α mutants in this study, and the chemical structure of 1NM-PP1. (B) Anti-Myc immunoblot of IRE1α transgenic proteins induced in INS-1 stable lines with doxycycline (Dox). (C) Confocal fluorescence micrographs of INS-1 cells showing co-localization of eGFP-WT IRE1α and eGFP-IRE1α (I642G) with transiently transfected ER-DsRED. Time course immunoblots of transgenic IRE1α proteins induced with Dox in INS-1 cells (anti-Myc and anti-phospho IRE1α). (D) Anti-total and anti-phospho IRE1α immunoblots of WT IRE1α, I642G, and K907A proteins produced from constructs transfected into Ire1 α−/− MEFs, +/− 1NM-PP1. (E) Immunoprecipitation (i.p) of Myc-tagged IRE1α proteins from T-REx 293 cells. (F) in vitro autophosphorylation of i.p.-ed Myc-tagged IRE1α and mutants, +/−1NM-PP1.