Appendix 1—key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) |
SIGMAR1 | GenBank | NM_005866 | |
| Gene (synthetic gene) | TM17 | GenScript | AAGAAATGG TGGGCTGCG GCGCTGTTA GCTGCGGCG TTACTGGCTG CGGCGTGGT GGAAGAAATAA |
|
| Gene (synthetic gene) | TM27 | GenScript | AAGAAATGG TGGGCTGCG GCGCTGTTAG CTGCGGCGTT ACTGGCTGCG GCGCTGCTTG CTGCGGCGTT ATTAGCTGCG GCGTGGTGGA AGAAATAA |
|
| Gene, fragment (Homo sapiens) |
IRE1a-TM | GenScript | GCGCGTCCGG AGGCGCCGGT GGACAGCATGC TGCAGGATATG GCGACCATCAT TCTGAGCACCT TCCTGCTGATC GGTTGGGTTGC GTTTATCATTAC CTACCCGCTGA GCAAG |
|
| Strain, strain background (Escherichia coli) | BL21(DE3) | Thermo Fisher | C600003 | Chemical competent cells |
| Cell line (Homo sapiens) | HEK293T | ATCC | 3216 | |
| Cell line (Homo sapiens) | HEK293 S1R KO | This study | Generated by co-transfection with Cas9 and guide plasmids, following by antibiotic selection with puromycin and blasticidin | |
| Biological sample (Mus musculus) | Liver | The Jackson Laboratory | 000664 | Freshly isolated from C57/B6 mouse strain |
| Antibody | Anti-phospho-IRE1a (rabbit polyclonal) | Novus Biologicals | NB100-2323 | WB (1:10,000) |
| Antibody | Anti-IRE1a (rabbit monoclonal) | Cell Signaling | 14C10 | WB (1:1000) |
| Antibody | Anti-XBP1s (rabbit monoclonal) | Cell Signaling | E9V3E | WB (1:1000) |
| Antibody | Anti-tubulin (mouse monoclonal) | DSHB | E7 | WB (1:500) |
| Antibody | Anti-APX2-HRP (rabbit polyclonal) | Abcam | ab192968 | WB (1:1000) |
| Antibody | Anti-S1R (mouse monoclonal) | Santa Cruz | B-5 | WB (1:300) |
| Antibody | Anti-His-tag (mouse monoclonal) | Millipore | HIS.H8 | WB (1:1000) |
| Antibody | Anti-STIM1 (rabbit polyclonal) | Cell Signaling | 4916 | WB (1:1000) |
| Antibody | Anti-calreticulin (rabbit polyclonal) | Abcam | Ab2907 | WB (1:5000) |
| Antibody | Anti-IP3R1 (rabbit polyclonal) | Home made | WB (1:1000) | |
| Antibody | Anti-TOM20 (rabbit monoclonal) | Cell Signaling | D8T4N | WB (1:1000) IF (1:250) |
| Antibody | Anti-GFP (chicken polyclonal) | Abcam | ab13970 | IF (1:1000–1:500) |
| Antibody | Anti-mCherry (rat monoclonal) | Invitrogen | 16D7 | IF (1:1000–1:500) |
| Antibody | Anti-TOM20 (rabbit polyclonal) | Santa Cruz | FL-145 | IF (1:500) |
| Antibody | Anti-mouse-HRP (goat polyclonal) | Jackson ImmunoResearch | 115-035-146 | WB (1:3000) |
| Antibody | Anti-rabbit-HRP (goat polyclonal) | Jackson ImmunoResearch | 115-035-144 | WB (1:3000) |
| Antibody | Anti-chicken-488 (goat polyclonal) | Invitrogen | A11039 | IF (1:1000) |
| Antibody | Anti-rat-594 (donkey polyclonal) | Invitrogen | A21209 | IF (1:1000) |
| Antibody | Anti-rabbin-Atto647N (goat polyclonal) | Sigma-Aldrich | 40839 | IF (1:1000) |
| Recombinant DNA reagent | pFast-Bac-HTA | Invitrogen | 10584027 | |
| Recombinant DNA reagent | Lentivector | Addgene | 14883 | |
| Recombinant DNA reagent | pEGFP-N2 | Clontech | PR29968 | |
| Recombinant DNA reagent | LentiGuide-Puro | Addgene | 52963 | |
| Recombinant DNA reagent | Lenti-Cas9-Blast | Addgene | 52962 | |
| Recombinant DNA reagent | S1R-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector | |
| Recombinant DNA reagent | S1R-R7ER8E-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced R7ER8E mutation | |
| Recombinant DNA reagent | S1R-ΔRR-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced ΔRR mutation | |
| Recombinant DNA reagent | S1R-4G-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced 4G mutation | |
| Recombinant DNA reagent | S1R-4A-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced 4A mutation | |
| Recombinant DNA reagent | S1R-W9L/W11L-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced W9L/W11L mutations | |
| Recombinant DNA reagent | S1R-Y173S-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced Y173S mutation | |
| Recombinant DNA reagent | S1R-Y201S/Y206S-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced Y201S/Y206S mutations | |
| Recombinant DNA reagent | S1R-Δ4-GFP | This study | Fusion gene of S1R and GFP in pEGFP-N2 vector with introduced Δ4 mutation | |
| Recombinant DNA reagent | mCherry-Sec61b | Addgene | 49155 | Zurek et al. Traffic. 2011 |
| Recombinant DNA reagent | mCherry-MAPPER | This study | mCherry-MAPPER was generated by replacing GFP with mCherry in the original construct from Chang et al. Cell Reports, 2013 | |
| Recombinant DNA reagent | His-S1R | This study | His-tagged S1R in pFastBac-HTA vector | |
| Recombinant DNA reagent | His-S1R-4G | This study | His-tagged S1R in pFastBac-HTA vector with introduced 4G mutation | |
| Recombinant DNA reagent | His-S1R-W9L/W11L | This study | His-tagged S1R in pFastBac-HTA vector with introduced W9L/W11L mutations | |
| Recombinant DNA reagent | S1R-6His | This study | His-tagged S1R in lentivector | |
| Recombinant DNA reagent | S1R-4G-6His | This study | His-tagged S1R in lentivector with introduced 4G mutation | |
| Recombinant DNA reagent | S1R-W9L/W11L-6His | This study | His-tagged S1R in lentivector with introduced W9L/W11L mutations | |
| Recombinant DNA reagent | MBP-TM17 | This study | Generated by cloning TM17 into pMAL-c5x vector | |
| Recombinant DNA reagent | MBP-TM27 | This study | Generated by cloning TM27 into pMAL-c5x vector | |
| Recombinant DNA reagent | MBP-IRE1a-TM | This study | Generated by cloning IRE1a-TM into pMAL-c5x vector | |
| Recombinant DNA reagent | S1R-APEX2 | This study | Generated by cloning S1R-APEX2 fusion gene into lentivector | |
| Recombinant DNA reagent | Sec61b-APEX2 | This study | Generated by cloning Sec61b-APEX fusion gene (RRID:Addgene_83411) into lentivector | |
| Recombinant DNA reagent | APEX2-KDEL | This study | Generated by adding KDEL-encoding sequence to the APEX2 gene and cloning into lentivector | |
| Sequence-based reagent | Guide sequence targeting exon 1 of human S1R | This study | GCGCGAAGAG ATAGCGCAGT |
|
| Sequence-based reagent | Guide sequence targeting LacZ | Platt et al. Cell, 2014 | GTGCGAATAC GCCCACGCGAT |
|
| Commercial assay or kit | Q5 site-directed mutagenesis kit | NEB | E0554 | |
| Chemical compound, drug | 18:1 PC (DOPC) | Avanti Polar Lipids | 850375 | |
| Chemical compound, drug | 24:1 PC (DNPC) | Avanti Polar Lipids | 850399 | |
| Chemical compound, drug | NBD-PC | Avanti Polar Lipids | 810130 | |
| Chemical compound, drug | Cholesterol | Avanti Polar Lipids | 700000 | |
| Chemical compound, drug | DSPE-PEG(2000)-biotin | Avanti Polar Lipids | 880129 | |
| Chemical compound, drug | DGS-NTA-Ni | Avanti Polar Lipids | 790404 | |
| Chemical compound, drug | Egg PC | Avanti Polar Lipids | 241601 | |
| Chemical compound, drug | LDAO | Anatrace | D360 | |
| Chemical compound, drug | OG | Anatrace | O311 | |
| Chemical compound, drug | DDM | Anatrace | D310 | |
| Chemical compound, drug | Cholesteryl hemisuccinate | Anatrace | CH210 | |
| Chemical compound, drug | CarboxyLink agarose | Thermo Scientific | 20266 | |
| Chemical compound, drug | Alexa647 NHS ester | Invitrogen | A37573 | |
| Chemical compound, drug | Alexa555 NHS ester | Invitrogen | A37571 | |
| Chemical compound, drug | C2 maleimide Alexa647 | Invitrogen | A20347 | |
| Chemical compound, drug | (+)-SKF-10047 | Sigma-Aldrich | A114 | |
| Chemical compound, drug | NE-100 hydrochloride | Tocris | 3133 | |
| Software, algorithm | MATLAB_R2019b | MATLAB | ||
| Software, algorithm | GraphPad Prism 8 | GraphPad | ||
| Other | Liposomal mini-extruder with 0.1 mkm polycarbonate filters | Avanti Polar Lipids | 610000 |