Peter van Mourik

73 Forskolin-induced swelling protocol KEY RESOURCES TABLE REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, Peptides and Recombinant Proteins 0.05% Trypsin EDTA Thermo Fisher Scientific : Invitrogen #25300-054 Advanced Dulbecco’s Modified Eagles Medium with Nutrient Mixture F-12 Hams (Ad-DF) 500 mL Thermo Fisher Scientific: Invitrogen #12634 B27 supplement Thermo Fisher Scientific: Invitrogen #17504-044 Calcein, AM Life Technologies: Gibco #C3100MP Dimethyl sulfoxide (DMSO) Sigma Aldrich #276855-1L Dulbecco’s modified Eagle medium (DMEM) Thermo Fisher Scientific: Invitrogen #31966-021 Fetal bovine serum (FBS, brand varies upon optimal batch selection) Sigma/ Bovogen Forskolin Sigma #F3917-10mg Gastrin Sigma Aldrich #G9145 Gentamicin Life Technologies: Gibco #15710-049 Glutamax Thermo Fisher Scientific: Invitrogen #35050 G418 (100 mg/mL) Invivogen #ant-gn-5 Human Epithelial Growth Factor (hEGF) Peprotech #AF-100-15 HEPES Thermo Fisher Scientific: Invitrogen #15630-056 Matrigel (important: protein concentration > 10 mg/mL) Corning #356231 N-Acetylcysteine Sigma Aldrich #A9165 Nicotinamide Sigma Aldrich #N0636 p38 MAPK inhibitor (p38i) (SB202190) Sigma Aldrich #S7067 Phosphate Buffered Saline 0 (without Ca and Mg) Sigma/ Thermofisher/ sterile homemade #D5652/ #14190250 Penicillin/ Streptomycin Thermo Fisher Scientific: Invitrogen #15140-122 Primocin (50 mg/ mL) InvivoGen #ant-pm-1 Recovery™ Cell Culture Freezing Medium Thermo Fisher Scientific: Invitrogen #12648010 Recombinant Human R-Spondin 3 Protein (hR-spondin-3) R&D #3500 – RS/ CF TGFb type I Receptor inhibitor (A83-01) Tocris #2939 Ultrapure 0.5 M EDTA, pH 8 Thermofischer #15575020 Vancomycin Sigma Aldrich #861987- 250mg VX-661 Selleckchem #S7059 VX-770 Selleckchem #S1144 4