Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Peptide, recombinant protein | Anx A1 | Abcam | ab86446 | |
| Peptide, recombinant protein | Anx A1 with N-terminal His tag | Abcam | ab184588 | |
| Peptide, recombinant protein | Anx A2 | Abcam | ab93005 | |
| Peptide, recombinant protein | Anx A6 | Abcam | ab92934 | |
| Peptide, recombinant protein | L2 peptide (GNRGTFIRGYKAMVMDMEFLYHVGYILTSVLGLFAHEL) | Peptide 2.0 | custom | |
| Peptide, recombinant protein | sL2 peptide (DVVIALHGNAMMYLLVFEHTSTGIGKFLRFYGERLMYG) | Peptide 2.0 | custom | |
| Peptide, recombinant protein | pL2 peptide (GNRGTFIRGYKAMVMDME) | Peptide 2.0 | custom | |
| peptide, recombinant protein | mpL2 peptide (K-Ahx-GNRGTFIRGYRAMVMDME, Ahx stands for 6-aminohexanoate residue) | Peptide 2.0 | custom | |
| Peptide, recombinant protein | smpL2 peptide (K-Ahx-RDYRGMRMIMGETFNVGA, Ahx stands for 6-aminohexanoate residue) | Peptide 2.0 | custom | |
| Peptide, recombinant protein | ANXA1 N-terminal peptide (MAMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPT) | Peptide 2.0 | custom | |
| Chemical compound, drug | siRNA buffer | Dharmacon | B-002000-UB-100 | |
| Chemical compound, drug | siRNA transfection reagent | Dharmacon | T-2001–02 | |
| Chemical compound, drug | diBrBAPTA (5,5′-dibromo1,2-bis(o-amino phenoxy)ehane -N,N,N′,N′-tetraacetic acid) | Invitrogen | D-1211 | |
| Chemical compound, drug | diBrBAPTA (5,5′-dibromo1,2-bis(o-amino phenoxy)ehane -N,N,N′,N′-tetraacetic acid) | Santa Cruz Biotechnology | sc-2273516 | |
| Chemical compound, drug | 2Hydroxyethyl)ethylenediaminetriacetic acid (HEDTA) | Sigma | H7154 | |
| Chemical compound, drug | inositol 1,4,5-trisphosphate | Invitrogen | I-3716 | |
| Chemical compound, drug | inositol 1,4,5-trisphosphate | Santa Cruz Biotechnology | sc-201521 | |
| Chemical compound, drug | NHS-activated magnetic beads | Pierce | 88826 | |
| Chemical compound, drug | Protein A Dynabeads | ThermoFisher | 10006D | |
| Chemical compound, drug | Anti-Flag M2 agarose beads | Sigma | A2220 | |
| Chemical compound, drug | Fura-2 AM | Molecular Probes | I-1225 | |
| Chemical compound, drug | siGLO Red transfection indicator | Dharmacon | D-001630-02-05 | |
| Chemical compound, drug | Cal-520/AM | AAT Bioquest | 21130 | |
| Chemical compound, drug | Caged Ins(1,4,5)P3/PM (caged InsP3) | Sirius Fine Chemical SiChem GmbH | cag-iso-2–145- | |
| Chemical compound, drug | EGTA/AM | ThemoFisher | E1219 | |
| Cell line (Gallus gallus) | DT40 cells (wild-type) | Riken Bioresource Center | RCB1464; RRID:CVCL_0249 | |
| Cell line (Gallus gallus) | DT40-KO cells (with all three InsP3R genes disrupted) | Riken Bioresource Center | RCB1467; RRID:CVCL_4634 |
|
| Cell line (Gallus gallus) | DT40-r3 cells | ref. (Mak et al., 2013b) in this study | NA | |
| Cell line (Homo-sapiens) | HEK293 cells | ATCC | CRL-1573; RRID:CVCL_0045
|
|
| Cell line (Homo-sapiens) | HEK-3KO cells | Kerafast | EUR030; RRID:CVCL_HB82 | |
| Cell line (Homo-sapiens) | HEK293-3KO-r InsP3R-3 cells | this study | NA | |
| Cell line Mus musculus | N2a cells | ATCC | CCL-131; RRID:CVCL_0470 | |
| Cell line (Rattus rattus) | PC12 cells | ATCC | CRL-1721; RRID:CVCL_0481 | |
| Cell line (Homo-sapiens) | tsA201 cells | Sigma-Aldrich | 96121229; RRID:CVCL_2737 | |
| Cell line (Homo-sapiens) | A549 cells | ATCC | CCL-185; RRID:CVCL_0023 | |
| Genetic reagent (Homo sapiens) | Anx A1 siRNA | Dharmacon | M-011161-01-0005 | |
| Genetic reagent (Homo sapiens) | Non-targeting siRNA | Dharmacon | D-001206-13-05 | |
| Antibody | rabbit polyclonal anti-AnxA1 antibody | Proteintech | 21990–1-AP; RRID:AB_11182596
|
WB: 1:1000-1:4000 IP: 1:1000-1:10000 IHC: 1:50-1:500 IF: 1:20-1:200 |
| Antibody | mouse monoclonal anti-AnxA1 antibody | ECM Biosciences | AM0211 | ELISA 1:1000 ICC 1:100 IP 1:100 WB 1:1000 |
| Antibody | rabbit polyclonal anti-FLAG antibody | Cell Signaling | 14793S; RRID:AB_2572291 |
WB: 1:1000 IP: 1:50 IHC: 1:800 IF: 1:800 FC: 1:1600 Chromatin IP: 1:50 |
| Antibody | goat anti-rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568 | Invitrogen | A-11011; RRID:AB_143157 | FC: 1–10 µg/mL ICC: 2 µg/mL IF: 2 µg/mL |
| Antibody | mouse monoclonal anti-calnexin antibody | Chemicon | MAB3126 RRID:AB_143157 |
ICC: 1:100-1:250 WB: 1:200-1:2000 IP: 1:200-1:1000 |
| Antibody | rabbit polyclonal anti-β actin antibody | Cell Signaling | 7881S; RRID:AB_1549731 |
capture Elisa: 1:100 |
| Antibody | goat polyclonal anti-mouse IgG-HRP antibody | Cell Signaling | 7074S; RRID:AB_2099233 |
capture Elisa: 1:1000-1:3000 |
| Antibody | horse polyclonal anti-mouse IgG-HRP antibldy | Cell Signaling | 7076S; RRID:AB_330924 | capture Elisa: 1:1000-1:3000 |
| Antibody | mouse monoclonal anti-βactin antibody | Cell Signaling | 8H10D10; RRID:AB_2242334
|
WB: 1:1000 IHC: 1:8000-1:32000 IF: 1:2500-1:10000 FC: 1:200-1:800 |
| Antibody | mouse monoclonal anti-type 3 InsP3R antibody | BD Transduction Laboratories | 610312; RRID:AB_397704 | WB: 1:2000-1:4000 |
| Software, algorithm | QuB | refs. (Qin et al., 2000) and (Bruno et al., 2013) in this study | Quantitative single-channel analysis | |
| Software, algorithm | IGOR Pro | Wavemetrics | Figure production and data fitting | |
| Software, algorithm | Metamorph v7.7 | Universal Imaging/Molecular Devices | Image analysis | |
| Software, algorithm | Flika | Ellefsen et al., 2014 | Image processing | |
| Software, algorithm | Microcal Origin v6.0 | OriginLab | Data analysis and graphing | |
| Software, algorithm | Max Chelator | online freeware | Calculation of ion concentrations | |
| Software, algorithm | MaxQuant, version 1.6.1.0 | online freeware | Database search |