Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | FRMD8; iTAP, human | n/a | ENSG00000126391 | |
| Gene (Mus musculus) | Frmd8; iTAP, mouse | n/a | ENSMUSG00000024816 | |
| Strain, strain background (Mm; C57BL/6) |
wild type; WT mice | Jackson Laboratories | RRID:IMSR_JAX:000664 | |
| Genetic reagent (Mm,C57BL/6) |
iTAP KO mice | this paper | n/a | Generated by CRISPR (more details in M and M section and Figure 7—figure supplement 1) |
| Cell line (Hs) | HEK 293ET | 10.1038/sj.emboj.7601743 | RRID:CVCL_6996 | |
| Cell line (Hs) | HeLa | ATCC | ATCC CCL-2; RRID:CVCL_0030 |
|
| Cell line (Mm) | RAW 264.7 | Sigma | 91062702; RRID:CVCL_0493 |
|
| Cell line (Mm) | L929 | ATCC | ATCC CCL-1; RRID:CVCL_0462 |
|
| Cell line (Hs) | HEK 293ET iTAP KO | this paper | n/a | Generated by CRISPR (more details in M and M section) |
| Cell line (Mm) | RAW 264.7 iTAP KO | this paper | n/a | Generated by CRISPR (more details in M and M section) |
| Cell line (Mm) | L929 iTAP KO | this paper | n/a | Generated by CRISPR (more details in M and M section) |
| Cell line (Mm) | MEF iTAP KO | this paper | n/a | Isolated from iTAP KO mouse embryos |
| Cell line (Mm) | DKO MEF | 10.1038/embor.2013.128. | n/a | Isolated from iRhom1/2 DKO mouse embryos |
| Biological sample (Mm) | tissues from WT and KO mice |
other | n/a | Isolated with standard techniques from the mice described here |
| Antibody | anti-TACE; TACE Ab 318 |
https://doi.org/10.1016
/j.jim.2011.06.015 |
n/a | |
| Antibody | anti-TACE for Immunoprecipitation |
R and D systems | R and D 9301 | |
| Antibody | anti-TACE | abcam | Ab39162; RRID:AB_722565 |
Ab39163 |
| Antibody | anti-P97 | Thermo-Fisher | MA1-21412; RRID:AB_557663 |
|
| Antibody | anti-TACE | R and D systems | 9301; RRID:AB_2223551 | |
| Antibody | anti-TACE | 10.1016/j.jim.2011.06.015 | Ab318 | monoclonal anti-TACE antibody (Trad et al., 2011) was used for the detection of deglycosylated TACE when no conA was used |
| Antibody | anti-tubulin | IGC antibody facility | Clone YL1/2; RRID:AB_793541 |
|
| Antibody | anti-HA-HRP | Roche | 3F10; RRID:AB_2314622 | |
| Antibody | anti-HA | Biolegend | 901501; RRID:AB_291259 | |
| Antibody | anti-V5-HRP | Life Technologies | R961-25 | |
| Antibody | anti-Frmd8 | Abnova | 157H00083786-B01P; RRID:AB_1573641 |
|
| Antibody | anti-Transferrin Receptor |
Life Technologies | 13–6800; RRID:AB_2533029 |
|
| Antibody | anti-Flag-HRP | Sigma | A8592; RRID:AB_439702 | |
| Antibody | anti-GAPDH | Cell signalling Technology | 2118; RRID:AB_561053 | |
| Antibody | anti-actin | Abcam | Ab8227; RRID:AB_2305186 | |
| Antibody | anti-iRhom2 | 10.1126/science.1214400 | n/a | Anti-iRhom2 polyclonal antibodies specific to the mouse iRhom2 N- terminus (amino acids 1–373) or raised against the iRhom homology domain, were previously described (Adrain et al., 2012) |
| Antibody | Anti-HA magnetic beads |
Pierce | 88836, Pierce | |
| Antibody | Mouse anti-GFP | IGC antibody Facility | clone 19F7 | Used as IP negative control in Figure 3—figure supplement 1 |
| Antibody | Anti-Flag M2 affinity gel |
Sigma | A220 | |
| Antibody | MagnaBind Goat Anti- Rabbit IgG Beads |
Thermo Scientific | 21356 | |
| Antibody | MagnaBind Goat Anti- Mouse IgG Beads |
Thermo Scientific | 21354 | |
| Antibody | mouse anti-LAMP2 | DSHB | Clone H4B4; RRID:AB_2134755 |
|
| Antibody | rabbit anti-EEA1 | Cell signalling Technology |
3288; RRID:AB_2096811 | |
| Antibody | rabbit anti-Strumpellin/ WASHC5 |
Santa Cruz | SC87442; RRID:AB_2234159 |
|
| Transfected construct (Hs) |
h iRhom1-HA (plasmid) | this paper | n/a | |
| Transfected construct (Hs) |
h iRhom1 Nterm HA (plasmid) |
this paper | n/a | |
| Transfected construct (Mm) |
Rhbdd2-HA (plasmid) | this paper | n/a | |
| Transfected construct (Hs) |
RHBDD3-HA (plasmid) | this paper | n/a | |
| Transfected construct (Mm) |
Ubac2-HA (plasmid) | this paper | n/a | |
| Recombinant DNA reagent |
pM6P.blast-GFP (plasmid) |
Felix Randow | n/a | |
| Recombinant DNA reagent |
pLEX.blast (plasmid) | this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
iRhom2-Cherry (plasmid) |
doi:10.1038/ni.3510 | n/a | |
| Transfected construct (Hs) |
ADAM33-V5 (plasmid) | 10.1038/embor.2013.128. | n/a | |
| Transfected construct (Hs) |
ADAM19-V5 (plasmid) | 10.1038/embor.2013.128. | n/a | |
| Transfected construct (Hs) |
ADAM22-V5 (plasmid) | 10.1038/embor.2013.128. | n/a | |
| Transfected construct (Hs) |
ADAM8-V5 (plasmid) | 10.1038/embor.2013.128. | n/a | |
| Transfected construct | TNF-FLAG (plasmid) | this paper | n/a | |
| Transfected construct (Hs) |
STING-FLAG (plasmid) | doi:10.1038/ni.3510 | n/a | |
| Transfected construct (Hs) |
LAMP1-mCherry (plasmid) |
this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
GFP-iRhom2 (plasmid) | this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
mCherry-iRhom2 (plasmid) |
this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
TACE-GFP (plasmid) | this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
TACE-TagRFP (plasmid) | this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
mouse iTAP-mCherry (plasmid) |
this paper | n/a | Details in M and M |
| Transfected construct (Hs) |
human iTAP-GFP (plasmid) |
this paper | n/a | Details in M and M |
| Transfected construct (Mm) |
iRhom2 NPAY > AAAA (plasmid) |
this paper | n/a | Quick change-based mutagenesis on iRhom2-HA plasmid described in M and M |
| Transfected construct (Mm) |
iRhom2 NRSY > AAAA (plasmid) |
this paper | n/a | Quick change-based mutagenesis on iRhom2-HA plasmid described in M and M |
| Transfected construct (Mm) |
iRhom2 Double NxxY > AAAA (plasmid) |
this paper | n/a | Quick change-based mutagenesis on iRhom2-HA plasmid described in M and M |
| Transfected construct (Hs) |
SREBP2-FLAG (plasmid) | DOI: 10.1083/jcb.201305076 | n/a | |
| Recombinant DNA reagent |
EGF-AP; EGF (plasmid) | doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
Betacellulin-AP; BTC-AP; BTC (plasmid) |
doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
TNF-AP; TNF (plasmid) | doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
TGFα-AP; TGFα (plasmid) | doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
HB-EGF-AP; HB-EGF (plasmid) |
doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
Epiregulin-AP; EPIREG-AP; EPIREG (plasmid) |
doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
Amphiregulin-AP; AREG-AP; AREG (plasmid) |
doi: 10.1083/jcb.200307137 | n/a | |
| Recombinant DNA reagent |
secreted luciferase | 10.1038/embor.2013.128. | n/a | |
| Recombinant DNA reagent |
pLenti-Crispr version2 (plasmid) |
Addgene | 52961 | |
| Recombinant DNA reagent |
pLentiCas9-Blast(plasmid) | Addgene | 52962 | |
| Recombinant DNA reagent |
pLentiGuide-Puro(plasmid) | Addgene | 52963 | |
| Recombinant DNA reagent |
pgRNAbasic(plasmid) | doi: 10.1242/dev.133074 | n/a | |
| Recombinant DNA reagent |
pT7-Cas9(plasmid) | doi: 10.1242/dev.133074 | n/a | |
| Sequence-based reagent (siRNA; Hs) |
iTAP oligo #1 | Santa Cruz | sc-96500 | |
| Sequence-based reagent (siRNA; Hs) |
iTAP oligo #2 | GE Dharmacon | M-018955-01-0005 | |
| Sequence-based reagent (siRNA; Hs) |
TACE oligo #1 | Santa Cruz | sc-36604 | |
| Sequence-based reagent (siRNA; Hs) |
TACE oligo #2 | GE Dharmacon | M-003453-01-0005 | |
| Sequence-based reagent (CRISPR Guide sequence; Hs; cell lines) |
gRNA targeting exon 1 | this paper | n/a | 5’-GCCCCGCTGAGCGATCCCAC-3’ |
| Sequence-based reagent (CRISPR Guide sequence; Hs; cell lines) |
gRNA targeting exon 4 | this paper | n/a | 5’-ACGTGTTCTTCCCAAAGCGG-3’ |
| Sequence-based reagent (CRISPR Guide sequence; Hs; cell lines) |
gRNA targeting exon 2 | this paper | n/a | 5'-TGACGTGCTGGTATACCTAG-3' |
| Sequence-based reagent (CRISPR Guide sequence; Hs; cell lines) |
gRNA targeting exon 6 | this paper | n/a | 5’-GGCACTTGAGGAGATAGGCG-3’ |
| Sequence-based reagent (CRISPR Guide sequence; Mm; cell lines) |
gRNA targeting first exon | this paper | n/a | 5’-TTCGGTGGGACCGCTCCGCA-3’ |
| Sequence-based reagent (CRISPR Guide sequence; Mm; cell lines) |
gRNA targeting second exon |
this paper | n/a | 5’-GCACTACTGTATCATCCGCC-3’ |
| Commercial assay or kit |
fluorogenic TACE substrate peptide |
ENZO Life Sciences | BML-P235-0001 | |
| Commercial assay or kit |
1-step PNPP Substrate | Thermo Fisher | PIER37621 | |
| Commercial assay or kit |
Fugene 6 | Promega | 2691-SC | |
| Commercial assay or kit |
Endoglycosidase; Endo-Hf | NEB | 174P0703 | |
| Commercial assay or kit |
PNGase F | NEB | 174P0704 | |
| Commercial assay or kit |
Concanavalin A Agarose; conA |
G-biosciences | 786–216 | |
| Commercial assay or kit |
MEGAshortscript T7 Kit | Thermo Fisher | AM1354 | |
| Commercial assay or kit |
MEGAclear kit | Thermo Fisher | AM1908 | |
| Commercial assay or kit |
Gibson Assembly Master Mix |
New England Biolabs | 174E2611 | |
| Commercial assay or kit |
KOD Hotstart DNA polymerase |
Novagen | 71086–5 | |
| Commercial assay or kit |
TOPO TA cloning kit for sequencing |
Invitrogen | 450030 | |
| Commercial assay or kit |
mMESSAGE mMACHINE T7 Ultra Kit |
Thermo Fisher | AM1345 | |
| Commercial assay or kit |
human TNF ELISA kit | R and D Systems | DY210 | |
| Commercial assay or kit |
human IL-6 ELISA kit | R and D systems | DY-206 | |
| Commercial assay or kit |
human IL-8 ELISA kit | R and D systems | DY-208 | |
| Chemical compound, drug |
Polyethylenimine; PEI | Sigma | 408727 | |
| Chemical compound, drug |
1,10-phenanthroline | Sigma | 131377 | |
| Chemical compound, drug |
Bafilomycin A1 | Santa cruz | 201550 | |
| Chemical compound, drug |
Chloroquine | Sigma | C6628 | |
| Chemical compound, drug |
Ammonium chloride | Acros | 10676052 | |
| Chemical compound, drug |
Ionomycin | Cayman | CAYM11932-5 | |
| Chemical compound, drug, (E.coli 055:B5) |
Lipopolysaccharide; LPS | Santa cruz | (sc-221855A) | |
| Chemical compound, drug |
Phorbol 12-myristate 13-acetate; PMA |
Sigma | P1585 | |
| Chemical compound, drug |
Cycloheximide; CHX | Santa cruz | sc-3508 | |
| Chemical compound, drug |
Dithiobis (succinimidyl propionate); DSP |
Pierce | 10731945 | |
| Chemical compound, drug |
Sulfo-NHS-LC-Biotin | Thermo Scientific, | 21335 | |
| Other | Lenti-X Concentrator | Clontech | Clonetech: 631231 | |
| Other | NeutrAvidin agarose; Neutravidin resin |
Thermo Fisher | 11885835 | |
| Other | NuPAGE Novex 4–12% Bit- Tris Protein Gels 1.0 mm |
Novex; Life Technologies | NP0322BOX | |
| Other | Opti-MEM I Reduced Serum Medium, GlutaMAX |
Life Technologies | 51985–026 | |
| Software, algorithm | Fiji | PMID: 22743772 | RRID:SCR_002285 | |
| Software, algorithm | Illustrator | Adobe | Adobe Creative Suite | |
| Software, algorithm | GraphPad Prism | GraphPad Software, Inc. | ||
| Software, algorithm | Volocity | PerkinElmer | ||
| Software, algorithm | Geneious | Biomatters Ltd. | Javaversion 1.8. 0_71-b15 |
|
| Software, algorithm | perkin elmer 2030 workstation |
PerkinElmer |