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. Author manuscript; available in PMC: 2015 Sep 4.
Published in final edited form as: Mol Cell. 2014 Jul 31;55(5):758–770. doi: 10.1016/j.molcel.2014.06.032

Figure 6. RtcB is in complex with IRE1α.

Figure 6

A. In HEK293T cells stably transfected with either Flag-RtcBWT or Flag-RtcBC122A, the level of Flag-RtcB was comparable to that of endogenous RtcB (lower panel, input). Anti-Flag co-IP showed that both wild type and ligase-dead RtcB proteins were associated with overexpressed IRE1α. Parental HEK293T cells that overexpressed IRE1α were used as a negative control. B. Using glutathione-Sepharose pull-down, a similar experiment was performed with GST-RtcB stably transfected HEK293T cells, showing that GST-RtcB interacted with overexpressed IRE1α. C. The association of RtcB with IRE1α was resistant to RNase A treatment. D–E. Endogenous IRE1α was associated with Flag-tagged or GST-tagged RtcB in two stable HEK293T cell lines that expressed Flag-RtcB and GST-RtcB, respectively.