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Journal Articles
The Journal of Cell Biology (2020) 219 (2): e201904048.
Published: 27 January 2020
Includes: Supplementary data
Images
Cells expressing IRE1β exhibit attenuated UPR signaling.  (A)  Tg-induced m...
Published: 27 January 2020
Figure 1. Cells expressing IRE1β exhibit attenuated UPR signaling. (A) Tg-induced mRNA expression of UPR genes assayed by qPCR for polarized Caco2 and T84 monolayers (log 2 [Tg-treated/DMSO control], n = 3). (B) Subtilase cytotoxin-induced mRNA expression of UPR genes assayed by qPCR for I... More
Images
Expression of IRE1β in intestinal epithelial cells and cell models.  Suppor...
Published: 27 January 2020
Figure S1. Expression of IRE1β in intestinal epithelial cells and cell models. Supports Fig. 1 . (A) Violin plots for expression of IRE1β and IRE1α transcripts in transcriptionally defined cell populations from mouse small intestine. Lines indicated median values. Data are from GSE92332 (Habe... More
Images
IRE1β suppresses IRE1  α  –XBP1 signaling.  (A and B)  XBP1 splicing was as...
Published: 27 January 2020
Figure 2. IRE1β suppresses IRE1 α –XBP1 signaling. (A and B) XBP1 splicing was assayed by (A) PCR and gel electrophoresis to detect spliced and unspliced XBP1 or (B) qPCR to detect spliced XBP1 transcript in HEK293 doxIRE1β cells treated with Dox and Tg as indicated ( n = 3). (C) Same as ... More
Images
IRE1β suppresses XBP1 transcriptional activity.  Supports  Fig. 2 .  (A)  H...
Published: 27 January 2020
Figure S2. IRE1β suppresses XBP1 transcriptional activity. Supports Fig. 2 . (A) HEK293T cells were cotransfected with UPRE-luciferase reporter and indicated plasmids, treated with Tg for 8 h, and assayed for luciferase activity. Activity is plotted relative to mock-transfected cells without ... More
Images
Suppression of IRE1α expression does not explain how IRE1β affects the UPR....
Published: 27 January 2020
Figure 3. Suppression of IRE1α expression does not explain how IRE1β affects the UPR. (A – C) IRE1α expression was assayed by (A) qPCR for mRNA or (B) immunoblot for protein in HEK293 doxIRE1β cells treated with Tg for indicated time points (immunoblots are representative of two or three ind... More
Images
IRE1β restricts stress-induced IRE1α expression.  Supports  Fig. 3 . Top: I...
Published: 27 January 2020
Figure S3. IRE1β restricts stress-induced IRE1α expression. Supports Fig. 3 . Top: IRE1α protein detected by immunoblot for HEK293T cells transiently transfected with control plasmid or IRE1β expression plasmid and treated with Tg for indicated time points. Bottom: Actin-normalized band intensi... More
Images
IRE1β enzymatic activity is not required to suppress stress-induced XBP1 sp...
Published: 27 January 2020
Figure 4. IRE1β enzymatic activity is not required to suppress stress-induced XBP1 splicing. (A) Schematic of IRE1β constructs. All constructs have C-terminal Myc tag. Expression of constructs in HEK293T cells was assayed by immunoblot with anti-Myc antibody. (B and C) XBP1 splicing activity ... More
Images
IRE1β interacts with IRE1α.  (A)  IRE1α and IRE1β proteins were assayed by ...
Published: 27 January 2020
Figure 5. IRE1β interacts with IRE1α. (A) IRE1α and IRE1β proteins were assayed by immunoblot with anti-IRE1α or anti-FLAG antibodies, respectively, for lysates or samples immunoprecipitated with anti-FLAG from HEK293 doxIRE1β cells treated with Dox and Tg as indicated. Blots are representativ... More
Images
IRE1β interacts with IRE1α.  Supports  Fig. 5 . IRE1α and IRE1β protein det...
Published: 27 January 2020
Figure S4. IRE1β interacts with IRE1α. Supports Fig. 5 . IRE1α and IRE1β protein detected by immunoblot of anti-FLAG immunoprecipitated (IP) samples or total lysates from HEK293 or HEK293/IRE1α KO cells transfected with IRE1β-FLAG expression construct. Endogenous IRE1α was detected with anti-I... More
Images
IRE1β inhibits IRE1α endonuclease activity in vitro.  (A)  Samples of affin...
Published: 27 January 2020
Figure 6. IRE1β inhibits IRE1α endonuclease activity in vitro. (A) Samples of affinity-purified full-length IRE1α and IRE1β were separated on SDS-PAGE and stained with Coomassie blue or assayed by immunoblot with anti-IRE1α, anti-IRE1β, or anti-Myc antibodies. (B) In vitro endonuclease activi... More
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Expression and XBP1 splicing activity of IRE1 constructs.  Supports  Fig. 6...
Published: 27 January 2020
Figure S5. Expression and XBP1 splicing activity of IRE1 constructs. Supports Fig. 6 . (A) IRE1α protein was assayed by immunoblot (top), spliced and unspliced XBP1 transcript was assayed by PCR and gel electrophoresis (middle), and spliced XBP1 transcript was assayed by qPCR (bottom) for HEK... More
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IRE1β has impaired phosphorylation and does not form higher-order oligomers...
Published: 27 January 2020
Figure 7. IRE1β has impaired phosphorylation and does not form higher-order oligomers. (A) XBP1 splicing luciferase reporter activity measured for HEK293T cells cotransfected with reporter and control vector or IRE1α(K599A) expression vector. Bars and error bars represent mean values ± SEM; sig... More
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A nonconserved amino acid in the kinase domain active site regulates IRE1 e...
Published: 27 January 2020
Figure 8. A nonconserved amino acid in the kinase domain active site regulates IRE1 endonuclease activity. (A) Top: Sequence alignment for IRE1α and IRE1β active site and activation loop residues. Conserved sequences are colored yellow (identical) and light yellow (similar). Bottom, left: Ribbo... More
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IRE1β-G641H mutant rescues XBP1 splicing.  Supports  Fig. 8 B . Spliced XBP...
Published: 27 January 2020
Figure S6. IRE1β-G641H mutant rescues XBP1 splicing. Supports Fig. 8 B . Spliced XBP1 transcript assayed by qPCR for HEK293/IRE1α KO cells transfected with indicated IRE1 constructs and treated with Tg. Bars represent a single independent experiment. These data were not included in Fig. 8 B ... More
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IRE1β functions as a dominant-negative suppressor of IRE1α signaling.  Sche...
Published: 27 January 2020
Figure 9. IRE1β functions as a dominant-negative suppressor of IRE1α signaling. Schematic illustrates model for IRE1β interacting with IRE1α oligomers in a manner to suppress stress-induced XBP1 splicing. More
Journal Articles
The Journal of Cell Biology (2020) 219 (2): e201911058.
Published: 24 January 2020
Images
Three discrete actin networks collaborate to create distinct functional reg...
Published: 24 January 2020
Figure 1. Three discrete actin networks collaborate to create distinct functional regions within the canonical “bullseye” IS. The outer ring of the IS (distal supramolecular activation cluster [dSMAC]) corresponds to the lamellipodium of a migrating cell. This region contains a prominent branche... More
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TCR deformation translates mechanical forces into biochemical signals  .  (...
Published: 24 January 2020
Figure 2. TCR deformation translates mechanical forces into biochemical signals . (A) In the resting state, ITAM motifs within the cytoplasmic domains of CD3 molecules interact with the inner leaflet of the T cell plasma membrane. (B) Protrusive forces applied by the T cell bring the TCR int... More
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Force contributes to T cell activation through several distinct actin struc...
Published: 24 January 2020
Figure 3. Force contributes to T cell activation through several distinct actin structures. (A) Overview of a T cell scanning an APC by migrating on its surface. During this process, discrete actin structures apply force at different areas of the T cell–APC interaction, facilitating T cell acti... More