Key resources table.
| Reagent type (species) or resource |
Designation | Sourcor reference | Identifiers | Additional Information |
|---|---|---|---|---|
| Danio Rerio casper strain (roy-/-; mitfa-/-) | PMID: 18371439 | |||
| Mus Musculus C.B-17/IcrHanTMHSD | Harlan, Indianapolis, IN, USA | |||
| Human primary juvenile foreskin lymphatic endothelial cells | Promocell, Heidelberg, Germany | |||
| Adult Dermal lymphatic endothelial cells | Lonza,Basel, Switzerland | |||
| WM852 | Wistar Institute Philadelphia | RRID:CVCL_6804 | ||
| WM165 | Wistar Institute Philadelphia | RRID:CVCL_L033 | ||
| Bowes | D.B. Rifkin, Rockfeller University | RRID:CVCL_3317 | ||
| WM793 | Wistar Institute Philadelphia | RRID:CVCL_8787 | ||
| HEK293FT | Biomedicum Functional Genomic Unit, University of Helsinki | |||
| siRNA | Notch3 siRNA Notch3 siRNA |
Dharmacon, Lafayette, CO Ambion,Waltham, MA |
L-011093-00-0005 4392420 |
|
| siRNA | MMP14 siRNA | Qiagen,Hilden, Germany | SI03648841; SI00071176 | |
| Transfected construct | NICD3pCLE | AddGene,Cambridge, MA;PMID: 16508304 | Plasmid #26894 | |
| Transfected construct | MMP14 Sport6 | GenomeBiology UnitUniversity of Helsinki |
NA | |
| Transfected construct | PcDNA3 | Invitrogen,Carlsbad, CA | NA | |
| Antibody | Pecam | DAKO,Santa Clara, CA | M0823 | IFA 3D 1:500 |
| Antibody | VE-cadherin | BD Pharmingen, San Jose, CA | 553927 | IFA 1:1000 |
| Antibody | GFP | Prof. Jason Mercer, UCL, London | NA | IFA 3D 1:1000 |
| Antibody | Notch3 | Santa Cruz Biotechnology,Dallas, TX |
sc-5593 | IFA 1:50 WB 1:250 |
| Antibody | MMP14 EP1264Y | Abcam,Cambridge, UK | 51074 | IFA 1:100 |
| Antibody | MMP14 (LEM clone) | Chemicom,Waltham, MA | MAB3328 | IFA 1:300 FACS 1:100 |
| Antibody | active β1-integrin 12G10 | Abcam | 30394 | IFA 1:300 FACS 1:100 |
| Antibody | active β1-integrin 9EG7 | BD Pharmingen | 553715 | IFA 1:100 FACS 1:100 |
| Antibody | Total β1-integrin | Abcam | 52971 | WB 1:1000 |
| Antibody | TGN46 | Sigma,St. Louis, MO | T7576 | IFA 1:500 |
| Antibody | LYVE1 | Reliatech,Wolfenbüttel, Germany | 103-PA50AG | IHC 1:200 |
| Antibody | Alexa594-phalloidin | Thermo Fisher,Waltham, MA | 21833 | IFA 3D 1:200 |
| Sequence-based reagents | ||||
| CD31 RTqPCR primers (for, rev) | AACAGTGTTGACATGAAGAGCC, TGTAAAACAGCACGTCATCCTT |
|||
| CD34 RTqPCR primers (for, rev) | TGGGCATCACTGGCTATTTC, CCACGTGTTGTCTTGCTGAA |
|||
| FLT4 RTqPCR primers (for, rev) | GACAGCTACAAATACGAGCATCTG, CTGTCTTGCAGTCGAGCAGAA |
|||
| NOTCH1 RTqPCR primers (for, rev) | GAGGCGTGGCAGACTATCATGC, CTTGTACTCCGTCAGCGTGA |
|||
| NOTCH2 RTqPCR primers (for, rev) | CCTGGGCTATACTGGGAGCTACTG ,ACACCCTGATAGCCTGGGACAC |
|||
| NOTCH4 RTqPCR primers (for, rev) | AATCCCACTGCCTCCAGACT, TTGTGGCAAAGGGAAGAGAC |
|||
| HES1 RTqPCR primers (for, rev) | TCAACACGACACCGGATAAA, TCAGCTGGCTCAGACTTTCA |
|||
| HEY1 RTqPCR primers (for, rev) | GTTCGGCTCTAGGTTCCATGT, CGTCGGCGCTTCTCAATTATTC |
|||
| HEY2 RTqPCR primers (for, rev) | TTGAGAAGACTTGTGCCAACTG, GTGCGTCAAAGTAGCCTTTACC |
|||
| MMP14 RTqPCR primers (for, rev) | GCAGAAGTTTTACGGCTTGCAA, CCTTCGAACATTGGCCTTGAT |
|||
| ACT RTqPCR primers (for, rev) | TCACCCACACTGTGCCATCTACGA, CAGCGGAACCGCTCATTGCCAATGG |
|||
| GAPDH RTqPCR primers (for, rev) | TCACCACCATGGAGAAGGCT, GCCATCCACAGTCTTCTGGG |
|||
| Commercial assay or kit | NucleoSpin RNA II kit | Macherey Nagel,Düren, Germany | 740955 | |
| Commercial assay or kit | SYBR Green PCR mix | Fermentas, Waltham, MA | 4415440 | |
| Commercial assay or kit | QuantiTect Primer Assay NOTCH3 | Qiagen | QT00003374 | |
| Commercial assay or kit | dextran coated magnetic nanoparticles extran coated magnetic nanoparticles | fluidMAG-DX, Chemicell, Berlin, Germany | 4104–5 | |
| Commercial assay or kit | MidiMACS separator | Miltenyi Biotec,Bergisch Gladbach, Germany | 130-042-302 | |
| Commercial assay or kit | LS column | Miltenyi Biotec | 130-042-401 | |
| Chemical compound, drug | Lipofectamine RNAiMax | Invitrogen | 13778150 | |
| Chemical compound, drug | DAPT | Sigma | D5942 | Used at 10 μM |
| Chemical compound, drug | GM6001 | Tocris Biosciences, Bristol, UK | 2983/10 | Used at 10 μM |
| Chemical compound, drug | NSC 405020 | Selleckchem,Munich, Germany | S8072 | Used at 50 μM |
| Chemical compound, drug | AIIB2 | DSHB hybridoma, from Johanna Ivaska | RRID:AB_528306 | Used 1:10 |
| Software, algorithm | Bioimage XD (http://www.bioimagexd.net/) | PMID: 22743773 | NA | |
| Software, algorithm | CellProfiler | PMID: 17076895 | RRID:SCR_007358 | |
| Software, algorithm | Adobe Photoshop | RRID:SCR_014199 | ||
| Software, algorithm | ImageJ | RRID:SCR_003070 | ||
| Software, algorithm | Pathview https://pathview.uncc.edu | RRID: SCR_002732 | ||
| Software, algorithm | Morpheus https://software.broadinstitute.org/morpheus/ | RRID: SCR_014975 | ||