Table 3.
Materials Table
| Reagent or resource | Source | Identifier |
|---|---|---|
| Antibodies | ||
| Alexa Fluor 700 anti-mouse CD45 | BioLegend | Cat# 157210; RRID: AB_2860730 |
| Alexa Fluor 700 anti-mouse Ly6G | BioLegend | Cat# 127621; RRID: AB_10640452 |
| APC anti-mouse Ly-6C | BioLegend | Cat# 128015; RRID: AB_1732087 |
| APC anti-mouse Ly-6G/Ly-6C (Gr-1) | BioLegend | Cat# 108411; RRID: AB_313376 |
| APC/Cyanine 7 anti-mouse CD326 (Ep-CAM) antibody | BioLegend | Cat# 118217; RRID: AB_1501158 |
| APC/Cyanine7 anti-mouse FcepsilonR1alpha antibody | BioLegend | Cat# 134325; RRID: AB_2572063 |
| Brilliant Violet 605 anti-mouse CD90.2 (Thy1.2) antibody | BioLegend | Cat# 140318; RRID: AB_2650924 |
| Brilliant Violet 785 anti-mouse CD117 (c-kit) | BioLegend | Cat# 105841; RRID: AB_2629799 |
| Goat polyclonal anti-rabbit horseradish peroxidase | Thermo Fisher | Cat# 32260; RRID: AB_1965959 |
| Goat polyclonal anti-rat AF 555 | Thermo Fisher | Cat# A-21434; RRID: AB_2535855 |
| Goat polyclonal anti-rabbit AF 555 | Thermo Fisher | Cat# A-21428; RRID: AB_2535849 |
| Pacific Blue anti-mouse CD45 antibody | BioLegend | Cat# 103126; RRID: AB_493535 |
| PE anti-mouse CD123 antibody | BioLegend | Cat# 106005; RRID: AB_2124403 |
| PE anti-mouse CD31 antibody | BioLegend | Cat# 102508; RRID: AB_312915 |
| PE-Cyanine 7 anti-mouse LYVE1 | Thermo Fisher | Cat# 25-0443-82; RRID: AB_2802237 |
| PE/Cyanine 7 anti-mouse/human CD11b | BioLegend | Cat# 101215; RRID: AB_312798 |
| PerCP/Cyanine 5.5 anti-mouse CD193 (CCR3) antibody | BioLegend | Cat# 144515; RRID: AB_2565741 |
| Rabbit monoclonal anti-Ki67 | Abcam | Cat# ab16667; RRID: AB_302459 |
| Rat monoclonal anti-BrdU | Abcam | Cat# ab6326; RRID: AB_305426 |
| Rat monoclonal anti-LYVE1 | Thermo Fisher | Cat# 14-0443-82; RRID: AB_1633414 |
| Rabbit polyclonal anti–COX-2 | Bioss | Cat# bs-0732R; RRID: AB_10859004 |
| Chemicals, peptides, and recombinant proteins | ||
| (2-Hydroxypropyl)-β-cyclodextrin | Sigma | Cat# 332593 |
| 10% neutral buffered formalin | VWR | Cat# 16004 |
| 16, 16-dimethyl prostaglandin E2 (dmPGE2) | Cayman Chemical Company | Cat# 14750 |
| 32% paraformaldehyde | Electron Microscopy Services | Cat# 15714 |
| 5-Bromo-2’-deoxyuridine (BrdU) | BioLegend | Cat# 423401 |
| Advanced Dulbecc’'s modified Eagle medium/F12 | Gibco | Cat# 12634-010 |
| Ampicillin | Sigma | Cat# A1593 |
| Antibiotic–antimycotic (anti-anti) | Thermo Fisher | Cat# 15240062 |
| B27 | Thermo Fisher | Cat# 17504044 |
| Bovine serum albumin | Sigma | Cat# A9418 |
| Cell staining buffer | BioLegend | Cat# 420201 |
| CHIR | Sigma | Cat# SML1046 |
| Collagenase II | Worthington | Cat# LS004176 |
| Cytodex 3 Microcarrier Beads | Cytiva | Cat# 17048502 |
| Dako Liquid DAB+ Substrate Chromogen System | Agilent Technologies | Cat# K3468 |
| DAPI | BioLegend | Cat# 422801 |
| DetachKit | Promocell | Cat# C-41200 |
| Dispase II | Sigma | Cat# D4693 |
| DNAse I | Sigma | Cat# 10104159001 |
| DPBS, no calcium, no magnesium | Gibco | Cat# 141190250 |
| DTT | Sigma | Cat# 43816 |
| EDTA | Thermo Fisher | Cat# 11575-038 |
| EGF | Thermo Fisher | Cat# PMG8043 |
| Endothelial Cell Growth Media MV2 | Promocell | Cat# C22121 |
| Fetal bovine serum | Gemini Bio-Products | Cat# 900-108 |
| Fibrinogen from bovine plasma, type I-S | Sigma | Cat# F8630 |
| Ficoll-Paque Premium | Cytiva | Cat# 17544202 |
| GFR Matrigel | Corning | Cat# 356231 |
| GlutaMAX | Gibco | Cat# 35050-061 |
| HBSS | Gibco | Cat# 14175-095 |
| HEPES | Sigma | Cat# H3662 |
| ImmPACT DAB Substrate Kit | Vector Laboratories | Cat# SK-4105 |
| L-161,982 (EP4i) | Cayman Chemical Company | Cat# 10011565 |
| LPS ultrapure | Invivogen | Cat# tlrl-3pelps |
| Metronidazole | Cayman Chemical Company | Cat# 9002409 |
| Molecular biology grade water | Corning | Cat# 46-000-CV |
| MSX-122 (CXCR4i) | Selleck Chemicals | Cat# S6617 |
| N-2 supplement | Thermo Fisher | Cat# 17502048 |
| N-acetyl-L-cysteine | Sigma | Cat# A9165 |
| Neomycin sulfate | Sigma | Cat# N6386 |
| Noggin | PeproTech | Cat# 250-38 |
| Nonessential amino acids | Thermo Fisher | Cat# 11140050 |
| Normal goat serum | Vector Laboratories | Cat# S-1000 |
| PerfeCTa SYBR Green FastMix, Low ROX | VWR | Cat# 101414-286 |
| Propidium iodide | BioLegend | Cat# 421301 |
| Qscript cDNA Supermix | VWR | Cat# 101414-108 |
| RBC lysis buffer (10X) | BioLegend | Cat# 420301 |
| RNAlater | Thermo Fisher | Cat# AM7020 |
| RPMI 1640 with L-glutamine | Thermo Fisher | Cat# 11875-119 |
| Rspo1 | R&D Systems | Cat# 3474-RS-050 |
| SB225002 (CXCR2i) | Selleck Chemicals | Cat# S7651 |
| SDS | Thermo Fisher | Cat# BP166-500 |
| Thrombin from bovine plasma | Sigma | Cat# T4648 |
| Triton X-100 | Thermo Fisher | Cat# BP151-500 |
| TRIzol LS | Thermo Fisher | Cat# 10296028 |
| TruStain FcX | BioLegend | Cat# 101319 |
| TWEEN 20 | Sigma | Cat# D8418 |
| TWEEN 80 | Sigma | Cat# P1754 |
| U0126 (MEK1/2i) | Selleck Chemicals | Cat# S1102 |
| Vancomycin | Sigma | Cat# SBR00001 |
| VECTASHIELD Antifade Mounting Medium with DAPI | Vector Laboratories | Cat# H-1200 |
| X-VIVO 15 media | Lonza | Cat# 04-418Q |
| Critical commercial assays | ||
| Human R-Spondin-3 Picokine ELISA Kit | Boster Bio | Cat# EK1512 |
| QIAmp DNA Stool Mini Kit | Qiagen | Cat# 51604 |
| Rneasy Mini Kit | Qiagen | Cat# 74104 |
| RT2 Profiler PCR Array | Qiagen | Cat# 009669 |
| Deposited data | ||
| Bulk RNAseq data of PGE2-treated HDLEC | This paper | GEO: GSE223854 |
| Experimental models: organisms/strains | ||
| Cell line: HDLEC (from juvenile foreskin) | Promocell | Cat# C-12216 |
| Mouse: HdcCre | The Jackson Laboratory | Cat# 021198 |
| Mouse: HdcCreERT2 | Chen et al,37 2017 | |
| Mouse: HdcGFP | Yang et al,38 2011 | |
| Mouse: Ptgs2fl/fl | Gift from Harvey Herschman, University of California Los Angeles | |
| Mouse: Rosa26-DTA | The Jackson Laboratory | Cat# 009669 |
| Software and algorithms | ||
| clusterProfiler | Wu et al,70 2021 | https://doi.org/10.18129/B9.bioc.clusterProfiler |
| FlowJo V10.8 | FlowJo, BD | https://www.flowjo.com |
| GraphPad Prism 9 | GraphPad Software | http://graphpad.com |
| ImageJ version 2.3/FIJI | Schindelin et al71, 2012 | http://imagej.net |
| QuPath V0.4 | Bankhead et al,66 2017 | http://qupath.github.io |
CCR3, C-C motif chemokine receptor 3; DAB, 3,3'-Diaminobenzidine; DAPI, 4′,6-diamidino-2-phenylindole; DPBS, Dulbecco's Phosphate-Buffered Saline; DTT, dithiothreitol; EGF, epidermal growth factor; ELISA, enzyme-linked immunosorbent assay; Ep-CAM, epithelial cellular adhesion molecule; MEK1/2i, mitogen-activated kinase kinase 1/2; PE, phycoerythrin; Ptgs2fl/fl, prostaglandin-endoperoxidase synthase 2 flox/flox; Rspo, R-spondin; SDS, sodium dodecyl sulfate.