Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| strain, strain background (Mus.musculus) |
CD1 | Department of Experimental Medicine, Faculty of Health and Medical Sciences, Unviersity of Copenhagen. |
||
| genetic reagent (M.musculus) |
Syt-1 nul allele (gene symbol: syt1) |
Geppert et al., 1994 | PMID: 18308932 | |
| genetic reagent (M.musculus) |
Munc13-2 null allele (gene symbol: Unc13b) |
Varoqueaux et al., 2002 | PMID: 12070347 | |
| genetic reagent (M.musculus) |
CAPS1 null allele (gene symbol: Cadps) |
Speidel et al., 2005 | PMID: 15820695 | |
| genetic reagent (M.musculus) |
CAPS2 null allele (gene symbol: Cadps2) |
Jockusch et al., 2007 | PMID: 18022372 | |
| cell line (HEK 293T) | HEK 293T | ATCC | CRL-1573 | Experiments in Figure 2b |
| cell line (HEK 293T) | HEK 293T | A gift from Dr. Theres Schaub and Prof Victor Tarabykin, Institute of Cell Biology and Cell Biology, Charité Berlin |
Experiments Figure 3a | |
| cell line (COS-7) | COS-7 | ATCC | CRL-1651 | |
| cell line (tsA201) | tsA201 | Sigma-aldrich | Sigma-aldrich: 96121229 | |
| transfected lentiviral construct (p156rrl-pCMV- PLCδ4PH-EGFP) |
PLCδ4-PH-GFP | This paper | Local reference: 131 | plasmid with PLCδ4 received from Thomas F. J. Martin (Department of Biochemistry, University of Wisconsin) |
| transfected construct (pCMV-PLCδ1-PH-EGFP) |
PLCδ1-PH-GFP | Michael Krauss (Leibniz- Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany). |
||
| transfected construct (pCMV-PLCδ1-PH-RFP) |
PLCδ1-PH-RFP | Ken Mackie (The Gill Center for Biomolecular Science, Bloomington, Indiana) |
||
| transfected construct (pCMV-mcherry-INPP5E) |
mCh-INPP5E | Posor et al., 2013 | ||
| transfected construct (pCMV-mRFPruby- N1*Lifeact) |
lifeact-RFP | Geerd van den Bogaart (Radboud University Medical Center, Nijmegen, The Netherlands) |
PMID: 18536722 | |
| commercial assay or kit | QIAprep Spin Miniprep Kit |
Qiagen | ||
| commercial assay or kit | QIAquick Gel Extraction Kit |
Qiagen | ||
| commercial assay or kit | QIAquick PCR Purification Kit |
Qiagen | ||
| chemical compound, drug | DMSO | Sigma-aldrich | Sigma-aldrich: D8418 | |
| chemical compound, drug | Ascorbic aci | Sigma-aldrich | Sigma-aldrich: A5960 | |
| chemical compound, drug | CaCl2 | Sigma-aldrich | Sigma-aldrich: 499609 | |
| chemical compound, drug | CellMask | Invitrogen | Invitrogen: C10046 | |
| chemical compound, drug | CsOH | Sigma-aldrich | Sigma-aldrich 516988 | |
| chemical compound, drug | DMEM | Gibco/Thermo Fisher | Gibco/Thermo Fisher: 31966047 |
Experiments in Figure 3a |
| chemical compound, drug | DMEM | Lonza | Lonza: BE12-741F | Experiments in Figure 2b,c |
| Chemical compound, drug | HBSS | Gibco/Thermo Fisher | 14025–050 | |
| chemical compound, drug | caged DOG-PI(4,5)P2 | This paper | European Molecular Biology Laboratory (EMBL), Cell Biology and Biophysics Unit, Meyerhofstr. 1, 69117 Heidelberg, Germany. Att: Carsten Schultz (schultz@embl.de) |
|
| chemical compound, drug | EDTA | Sigma-aldrich | Sigma-aldrich: E5134 | |
| chemical compound, drug | Fetal Bovine Serum (FBS) |
Gibco/Thermo Fisher | Thermo Fisher/Gibco: 16140063 |
Experiments in Figure 3a |
| chemical compound, drug | Fetal Bovine Serum (FBS) |
Gibco/Thermo Fisher | Thermo Fisher/Gibco: 10270–106 |
Experiments in Figure 2b,c |
| chemical compound, drug | Fura-4F | Invitrogen | Invitrogen: F14174 | |
| chemical compound, drug | Furaptra | Invitrogen | Invitrogen: M1290 | |
| chemical compound, drug | Glucose | Sigma-aldrich | Sigma-aldrich: G8270 | |
| chemical compound, drug | HEPES | Sigma-aldrich | Sigma-aldrich: H3375 | |
| chemical compound, drug | Insulin-transferrin- selenium-X |
Invitrogen | Invitrogen: 51500056 | |
| chemical compound, drug | KCl | Sigma-aldrich | Sigma-aldrich: P5405 | |
| chemical compound, drug | L-Cysteine | Sigma-aldrich | Sigma-aldrich: C7352 | |
| chemical compound, drug | L-Glutamic acid | Sigma-aldrich | Sigma-aldrich: G1251 | |
| chemical compound, drug | Lipofectamin 2000 | Thermo Fisher | Thermo Fisher: 11668027 | |
| chemical compound, drug | Lipofectamin LTX | Thermo Fisher | Thermo Fisher: 15338100 | |
| chemical compound, drug | Opti-MEM I Reduced Serum Medium |
Thermo Fisher | Thermo Fisher: 31985070 | |
| chemical compound, drug | Dulbecco's Modified Eagle Medium |
Thermo Fisher | ThermoFirsher: 31966021 | |
| chemical compound, drug | Mg-ATP | Sigma-aldrich | Sigma-aldrich: A9187 | |
| chemical compound, drug | MgCl2 | Sigma-aldrich | Sigma-aldrich: 449172 | |
| chemical compound, drug | NaCl | Sigma-aldrich | Sigma-aldrich: S9888 | |
| chemical compound, drug | Na-GTP | Sigma-aldrich | Sigma-aldrich: G8877 | |
| chemical compound, drug | NaH2PO4 | Sigma-aldrich | Sigma-aldrich: S8282 | |
| chemical compound, drug | NPE | Synaptic Systems | SySy: 510 006 | |
| chemical compound, drug | Papain | Worthington Biochemical | Worthington Biochemical: LS003126 |
|
| chemical compound, drug | Penicillin/ streptomycin |
Invitrogen | Invitrogen: 15140122 | |
| chemical compound, drug | Pluronic F-127 | Thermo Fisher | Thermo Fisher: P3000MP | |
| chemical compound, drug | cg-DAG | Nadler et al., 2013 | PMID: 23720390 | |
| chemical compound, drug | caged SAG-PI(4,5)P2 | This paper | European Molecular Biology Laboratory (EMBL), Cell Biology and Biophysics Unit, Meyerhofstr. 1, 69117 Heidelberg, Germany. Att: Carsten Schultz (schultz@embl.de) |
|
| chemical compound, drug | trypsin-inhibitor | Sigma-aldrich | Sigma-aldrich: T9253 | |
| chemical compound, drug | U73122 | Sigma-aldrich | Sigma-aldrich: U6756 | |
| chemical compound, drug | U73343 | Sigma-aldrich | Sigma-aldrich: U6881 | |
| software, algorithm | Igor Pro | Wavemetrics | ||
| ImageJ version 1.50b | Waybe Rasband, National Institute of Health, USA |
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| SigmaPlot v. 12.3 | Systat Software Inc. | |||
| Matlab | MathWorks |