KEY RESOURCES TABLE.
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Human/Mouse Active Caspase-3 Antibody | R&D Systems | Cat#AF835 |
Mouse Integrin alpha 9 Antibody | R&D Systems | Cat#AF3827 |
SOX2 Monoclonal Antibody (Btjce), eBioscience™ | Invitrogen | Cat#14-9811-82; RRID: AB_11219471 |
LEF1 Monoclonal Antibody (E.281.8) | Invitrogen | Cat#MA5-14966; RRID: AB_10987030 |
Recombinant Anti-GATA3 antibody [EPR16651] - ChIP Grade | Abcam | Cat#ab199428 |
anti-acidic Hair Keratin K31 guinea pig polyclonal, serum | Progen | Cat#GP-hHa1 |
Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb | Cell Signaling | Cat#18338 |
Anti-Fibronectin Antibody | Sigma-Aldrich | Cat#AB2033 |
anti-acidic Hair Keratin K32 guinea pig polyclonal, serum | Progen | Cat#GP-HHA2 |
TCHH Polyclonal Antibody | Invitrogen | Cat#PA5-120392; RRID: AB_2913964 |
Anti-Ki67 antibody | Abcam | Cat#ab15580 |
Anti-alpha smooth muscle actin antibody | Abcam | Cat#ab5694 |
Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 | Invitrogen | Cat#A32731; RRID: AB_2633280 |
Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 | Invitrogen | Cat#A-11055; RRID: AB_2534102 |
Donkey anti-Rat IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 | Invitrogen | Cat#A-21208; RRID: AB_2535794 |
Goat anti-Guinea Pig IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 | Invitrogen | Cat#A-11073; RRID: AB_2534117 |
Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 633 | Invitrogen | Cat#A-21071; RRID: AB_2535732 |
Chemicals, Peptides, and Recombinant Proteins | ||
CRISPR-spCas9 nuclease | JHUSOM Transgenic Mouse Core Facility | RRID:SCR_017809 |
Fetal Bovine Serum (FBS) | Hyclone | Cat#SH30071.02HI |
Penicillin-Streptomycin | Sigma | Cat#P0781 |
Tamoxifen | Sigma | Cat#T5648-5G |
Corn oil | Sigma | Cat#C8267-2.5L |
GLYCEROL, ANHYDROUS “SUPER GLYCEROL” | AmericanBio | Cat#AB00751-00500 |
EDTA, 0.5M SOLUTION, PH 8.0 | AmericanBio | Cat#AB00502-01000 |
Human Plasma Fibronectin | EMD Millipore | Cat#FC010 |
TGF-β1 | R&D | Cat#7754-BH |
SlowFade Diamond Antifade Mountant | Invitrogen | Cat#S36972 |
Paraformaldehyde (PFA) 16% Aqueous SOL. EM GRADE | Electron Microscopy Sciences | Cat#15710 |
Aqueous Glutaraldehyde EM Grade, 10% 10x10ml | Electron Microscopy Sciences | Cat#16120 |
Sodium Cacodylate Buffer, 0.2M | Electron Microscopy Sciences | Cat#11652 (500ML) |
Osmium Tetroxide 4% solution | Electron Microscopy Sciences | Cat#19150 |
Toluidine Blue O | Electron Microscopy Sciences | Cat#22050 |
Uranyl Acetate | Electron Microscopy Sciences | Cat#22400 |
Lead Citrate | Fluka | Cat#15326 |
Critical Commercial Assays | ||
goat anti-rabbit IgG Alexa Fluor™ 488 Tyramide SuperBoost™ Kit | Invitrogen | Cat#B40922 |
SiR-DNA Kit | Cytoskeleton, Inc | Cat#SKU CY-SC007 |
PELCO® Eponate 12™ Kit with BDMA | Ted Pella, Inc. | Cat#18012 |
Experimental Models: Organisms/Strains | ||
Mouse: Pdgfrα-H2BGFP: B6.129S4-Pdgfratm11(EGFP)Sor/J | The Jackson Laboratory | RRID:IMSR_JAX:007669 |
Mouse: Pdgfrα-CreER: B6N.Cg-Tg(Pdgfra-cre/ERT)467Dbe/J | The Jackson Laboratory | RRID:IMSR_JAX:018280 |
Mouse: Rosa26-loxP-membraneTomato-(stop)-membraneGFP(mTmG): B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J | The Jackson Laboratory | RRID:IMSR_JAX:007676 |
Mouse: ROSA-LSL-DTA: B6.129P2-Gt(ROSA)26Sortm1(DTA)Lky/J | The Jackson Laboratory | RRID:IMSR_JAX:009669 |
Mouse: R26-LSL-tdTomato: B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J | The Jackson Laboratory | RRID:IMSR_JAX:007909 |
Mouse: Tgfbr1fl/fl: STOCK Tgfbr1tm1.1Kar/KulJ | V. Kaartinen (Larsson et al.68) | RRID:IMSR_JAX:028701 |
Mouse: B6SJL/F2 | JHUSOM Transgenic Mouse Core Facility | RRID:SCR_017809 |
Mouse: mNeonGreen-Smad4: B6SJL-Smad4tm1(mNG)+/− | This Paper | N/A |
Mouse: CD1: Crl:CD1(ICR) | Charles River | Strain #022 |
Oligonucleotides | ||
CRISPR gRNA: 5’ ACTATGTACAATGCTCAGAC 3’ | This Paper | N/A |
PCR primer for gBlock amplification, Forward: 5’ GCC TTT GCA ATA GTG AGG AGA CCG G | This Paper | N/A |
PCR primer for gBlock amplification, Reverse: 5’ GCA TCT GGG AAT GCT CTC TTC TCG CC | This Paper | N/A |
Recombinant DNA | ||
Linear dsDNA gBlock (837bp) HDR template: See Methods: Mice for sequence information |
This Paper | N/A |
Software and Algorithms | ||
ZEN 2.3 pro | ZEISS | micro-shop.zeiss.com/en/de/softwarefinder/software-categories/zen-blue/zen-blue-pro/ |
Fiji (ImageJ) | NIH | fiji.sc |
GraphPad Prism 9 | GraphPad Software, inc | graphpad.com |
Imaris | Bitplane/Oxford Instruments | imaris.oxinst.com |
Adobe Illustrator | Adobe | adobe.com |
Adobe Premiere Pro | Adobe | adobe.com |