Table of Materials
| Name of Reagent/Equipment | Company | Catalog Number | Comments/Description |
|---|---|---|---|
| 2-Amino-2-(hydroxymethyl)-1,3-propanediol | Sigma | T4661 | Tris base, Component of lysis buffer |
| A-83–01 | Tocris | 2939 | ALK4/5/7 inhibitor |
| Acetic acid, glacial | Fisher Scientific | A38 | |
| Advanced DMEM/F12 | Life Technologies | 12634–010 | Component of growth medium |
| Alexa Fluor 488 phalloidin | Life Technologies | A12379 | Fluorescent probe for F-actin |
| Antibiotic/antimycotic cocktail | Invivogen | ant-pm-2 | Primocin (100x) |
| B27, 50x | Life Technologies | 17504–044 | Component of growth medium |
| Cell culture inserts | Corning | 3470 | Transwell, PET membrane, 0.4 μm pore, 24-well plate |
| CHIR 99021 | Tocris | 4423 | GSK3β inhibitor |
| Click-iT Plus EdU Alexa Fluor 594 Imaging Kit | Life Technologies | C10639 | |
| Collagen IV, from human placenta | Sigma | C5533 | |
| Cultrex Organoid Harvesting Solution | Trevigen | 3700-100-01 | Depolymerizes basement membrane matrix |
| Enterohemorrhagic Escherichia coli (EHEC) | Kaper lab, University of Maryland | ||
| Epithelial voltohmmeter | World Precision Instruments | EVOM2 | |
| Epithelial voltohmmeter electrode | World Precision Instruments | STX3 | |
| Escherichia coli strain HS | Kaper lab, University of Maryland | ||
| Ethanol, absolute | Pharmco | 111000200 | |
| FluorSave mounting medium | Millipore | 345789 | For mounting insert membrane on microscope slide |
| GlutaMAX | Life Technologies | 35050–061 | L-alanyl-L-glutamine dipeptide, 200 mM |
| HEK293T/Noggin-Fc cell line | van den Brink lab, Tytgat Institute for Liver and Intestinal Research | For production of Noggin conditioned medium | |
| HEK293T/RSPO1-Fc-HA cell line | Trevigen | 3710–001-K | For production of Rspondin-1 conditioned medium |
| HEPES, 1 M | Life Technologies | 15630–080 | Component of growth medium |
| Heraeus Multifuge X1R Centrifuge | Thermo Fisher Scientific | 75004250 | |
| Hoechst 33342 | Life Technologies | H3570 | Fluorescent nuclear dye |
| Human epidermal growth factor (EGF) | R&D Systems | 236-EG-01M | Component of growth medium |
| IGEPAL CA-630 | Sigma | I8896 | Component of lysis buffer |
| Inverted cell culture light microscope | Olympus | CKX51 | |
| LB broth | EMD Millipore | 1.10285.0500 | |
| L-WNT3A cell line | ATCC | CRL-2647 | For production of Wnt3a conditioned medium |
| Matrigel, growth factor reduced | Corning | 356231 | Basement membrane matrix for 3D culture |
| Mini cell scraper | United BioSystems | MCS-200 | |
| MUC2 antibody, mouse monoclonal | Abcam | ab11197 | Use at 1:100 for immunostaining, 1:500 for immunoblotting |
| MUC2 shRNA lentiviral particles | GE Dharmacon | RHS4531 | GIPZ lentiviral shRNA, ID: 4583 |
| Orbital shaker | Grant Instruments | PSU-10i | |
| Penicillin-streptomycin, 100x | Life Technologies | 15140–122 | Component of growth medium |
| Phosphate buffered saline | Corning | 21–031 | |
| Probe sonicator with microtip | Branson Ultrasonics | 450 | |
| Protease inhibitor cocktail for mammalian cells | Sigma | P8340 | Component of lysis buffer |
| SB 202190 | Tocris | 1264 | p38 MAPK inhibitor |
| Sodium chloride | Sigma | S3014 | Component of lysis buffer |
| Sodium dexoycholate | Sigma | D6750 | Component of lysis buffer |
| Sodium dodecyl sulfate | Sigma | L3771 | Component of lysis buffer |
| TrypLE Express, 1x | Life Technologies | 12605–010 | Trypsin, for digesting enteroid/colonoid fragments |
| Vinculin antibody, rabbit monoclonal | Abcam | ab129002 | Use at 1:1000 for immunoblotting |
| Water, tissue culture grade, sterile filtered | Corning | 25–055 | |
| Y-27632 | Tocris | 1254 | RhoA/ROCK inhibitor |