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. 2015 May 18;35(12):2186–2202. doi: 10.1128/MCB.00013-15

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Copyright © 2015, Bright et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 Unported license.

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FIG 5 — BLOC1S1 is cleaved by IRE1 at guanine 444 in vitro. (A) Sequence of BLOC1S1 surrounding guanine 444 that was identified as a consensus IRE1 target sequence bioinformatically. (B) Predicted secondary structure of full-length BLOC1S1 mRNA folded in silico using the RNAfold Web server. The stem-loop surrounding G444 is magnified and annotated in panel A. The colors represent the probability of base pairing. (C) (Left) Urea gel electrophoresis of XBP1u(266–602), WT BLOC1S1, and G444C BLOC1S1 RNA after in vitro cleavage by the indicated concentrations of IRE1 for 15 min. (Right) Urea gel electrophoresis of the indicated RNAs after in vitro cleavage by IRE1 (0.5 μM) for the indicated times. The identities of RNA fragments are shown between the two gels; size markers are shown on the right. (D) Sequence of human BLOC1S1 from the start codon to the poly(A) tail. All occurrences of the sequence identical to the first 6 bases of the loop sequence around G444 are indicated in boldface.