KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
|
| ||
| Antibodies | ||
|
| ||
| Rabbit polyclonal anti-Ang1 | Abcam | ab8451 |
| anti-chicken-Alexa488 | Molecular Probes | A11039 |
| anti-goat-Alexa488 | Jackson ImmunoResearch | 705–545-033 |
| anti-goat-Alexa568 | Molecular Probes | A11057 |
| anti-mouse-Alexa488 | Jackson ImmunoResearch | 115–545-146 |
| anti-mouse-Alexa568 | Invitrogen | A11031 |
| anti-rabbit-Alexa488 | Invitrogen | A11008 |
| anti-rabbit-Alexa488 | Invitrogen | 711–545-152 |
| anti-rabbit-Alexa568 | Jackson ImmunoResearch | A10042 |
| anti-rat-Alexa594 | Molecular Probes | A21209 |
| Mouse monoclonal anti-CALBINDIN-D-28K(clone CB-955) | Sigma-Aldrich | C9848 |
| Rat monoclonal anti-CD13 | Bio-Rad | MCA2183GA |
| Goat polyclonal anti-CD31 | R&D | AF3628 |
| DAPI | Invitrogen | D1306 |
| Goat polyclonal anti-GFAP | Abcam | ab53554 |
| Chicken polyclonal anti-GFP | Aves | GFP-1020 |
| IsolectinB-Alex568 | Invitrogen | I21412 |
| Mouse monoclonal anti-OLIG2 | Millipore | MABN50 |
| TO-PRO-3 | Invitrogen | T3605 |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Agarose | Sigma-Aldrich | A9538 |
| Ampicillin sodium salt | Sigma-Aldrich | A9518 |
| Anti-HA-tag mAb-Magnetic Beads | MBL | M180–11 |
| Bovine Serum Albumin (BSA) | Roth | 8076.2 |
| Calcium chloride dihydrate | AppliChem | A3587 |
| cDNA synthesis SuperScript® Vilo | Invitrogen | 11754–050 |
| D(+)-Saccharose | Roth | 4621.1 |
| Deoxynucleotides (dNTPs) | NEB | N0447S |
| DEPC | Sigma-Aldrich | D5758 |
| Dextran Sulfate Sodium (DSS) | MP Biomedicals | SKU 0216011050 |
| DIG Blocking Reagent | Roche | 11096176001 |
| Digoxigenin (DIG) RNA labeling kit | Roche | 11175025910 |
| Dimethylsulfoxid (DMSO) | Gruesing | 10282 |
| DNA ladder 1 kb | Thermo Scientific | SM0311 |
| DNA ladder 100 bp | NEB | N3231S |
| DNA loading dye (6×) | Thermo Scientific | P0611 |
| DNA stain | Serva | 39803.01 |
| DNase I, Rnase-free | Thermo Scientific | EN0521 |
| DNaseI | Sigma-Aldrich | 4716728001 |
| EDTA | AppliChem | A3553 |
| Ethanol | Sigma-Aldrich | 32205 |
| Fast Green FCF | Sigma-Aldrich | F7252 |
| Fast SYBR Green Master Mix | Thermo Scientific | 408995 |
| Fluoromount-G | Linaris | 0100–01 |
| Formamid | Sigma-Aldrich | 47670 |
| Gelatine from porcin skin | Sigma-Aldrich | G1890 |
| GenElute Plasmid Miniprepkit | Sigma-Aldrich | PLN70–1Kt |
| Hydrochloric acid solution, 1 M | Sigma-Aldrich | 35328 |
| Isofluran | Sigma-Aldrich | 792632 |
| Isopropanol | VWR | ACRO423830010 |
| Ketamin (10%) | Bremer Pharma GmbH | 27015.00.00 |
| LB-Medium (BactoTMAgar) | BD | 214010 |
| LB-Medium (BactoTMTryptone) | BD | 211705 |
| LB-Medium (BactoTMYeast Extract) | BD | 212750 |
| Levamisol | Sigma-Aldrich | BP212 |
| Lidocain | Aspen Pharma Trading Limited | PL 39699/0086 |
| Magnesium chloride | AppliChem | A3618 |
| Maleic acid | Sigma-Aldrich | M0375 |
| mAng1 recombinant protein | R&D Systems | 9936-AN |
| mAng2 recombinant protein | R&D Systems | 7186-AN |
| Maxima Reverse Transcriptase | Thermo Scientific | EP0742 |
| Methanol | Sigma-Aldrich | 32213 |
| Mowiol® 4–88 | Sigma-Aldrich | 81381 |
| NBT/BCIP | Promega | S3771 |
| NEB T4 DNA Ligase | NEB | M0202 |
| NEG-50TM | Thermo Scientific | 6502 |
| Normal donkey serum | Dianova | 017–000-121 |
| Normal goat serum | Dianova | 005–000-121 |
| NucleoSpin® Gel and PCR Clean-Up | Nachery-Nagel | 740609.25 |
| PCR-CleanUp | Promega | A9281 |
| Phosphate Buffer Saline | Sigma-Aldrich | D8537 |
| Plasmid Plus MAXI kit | QIAGEN | 12963 |
| Potassium chloride | AppliChem | A3582 |
| Potassium dihydrogenphosphate | AppliChem | A3620 |
| Q5® HighFidelity DNA Polymerase | NEB | M0491 |
| Sodium bicarbonate | Sigma-Aldrich | 31437 |
| Sodium chloride | Sigma-Aldrich | 31434 |
| Sodium dodecyl sulfate (SDS) | Roth | CN30.3 |
| Sodium hydroxyde | Sigma-Aldrich | 30620 |
| Sodium phosphate dibasic dihydrate | Sigma-Aldrich | 4272 |
| Sodium phosphate monobasic monohydrate | Sigma-Aldrich | 71504 |
| Tissue-Tek® O.C.T. Compound | Genprice | 4583 |
| Tneasy® Mini kit | QIAGEN | 74104 |
| Torula RNA | Merck | 10109223001 |
| tris(hydroxymethyl)aminoethane (TRIS) | Roth | 4855.2 |
| Triton X-100 | Merck | 108603 |
| TRIzol Reagent | Thermo Scientific | 15596026 |
| Trypsine-EDTA | Sigma-Aldrich | T3924 |
| Tween®20 | Roth | 9127.1 |
| Wizard®SV Gel and PCR CleanUp System | Promega | A9281 |
| Xylavet (20 mg/ml) | CP-Pharma Handelsges. mbH | 401510.00.00 |
|
| ||
| Deposited data | ||
|
| ||
| RNA-seq data | this study | GSE178659 |
|
| ||
| Experimental models: Organisms/strains | ||
|
| ||
| Ang1flox/flox | (Jeansson et al., 2011) | N/A |
| Ang2−/− | (Hu et al., 2014) | N/A |
| NesCre | (Tronche et al., 1999) | N/A |
| Rosa26:mTomato-mGFP | (Muzumdar et al., 2007) | N/A |
| Jackson Laboratories | ||
| JAX stock #007676 | ||
| Pcp2Cre | (Barski et al., 2000) | N/A |
| Rpl22HA/HA | (Sanz et al., 2009) | N/A |
| Jackson Laboratories | ||
| JAX stock #011029 | ||
| Tie2flox/flox | (Savant et al., 2015) | N/A |
| Tie2-GFP | (De Palma et al., 2005) | N/A |
|
| ||
| Oligonucleotides | ||
|
| ||
| See Table S3 | N/A | |
|
| ||
| Software and algorithms | ||
|
| ||
| DEseq2 | (Love et al., 2014) | https://bioconductor.org/packages/release/bioc/html/DESeq2.html |
| FastQC | Freeware (Andrews, 2010) | https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ |
| Fiji - ImageJ | Freeware (Schindelin et al., 2012) | https://imagej.net/software/fiji |
| GraphPad Prism 6/7 | GraphPad Software | https://www.graphpad.com |
| HTSeq-count | (Anders et al., 2015) | https://htseq.readthedocs.io/en/release_0.11.1/overview.html |
| IMARIS 8.4.2 | Bitplane AG | https://imaris.oxinst.com/ |
| IPA | QIAGEN Inc. (Krämer et al., 2014) | https://digitalinsights.qiagen.com/products |
| Microsoft Office | Microsoft | https://office.live.com/start/Excel.aspx |
| R | Freeware (R Team, 2020) | https://cran.r-project.org/ |
| samtools | (Li et al., 2009) | http://www.htslib.org/ |
| STAR | (Dobin et al., 2013) | https://github.com/alexdobin/STAR |