Rabbit antibodies against

Fetal bovine serum (FBS), trypsin, penicillin, streptomycin, and Dulbecco’s modified Eagle’s medium (DMEM) were from Gibco BRL (Grand Island, NY, USA). Rabbit antibodies against moesin phosphorylated on Thr558, desmin, and CD44 were from Abcam (Cambridge, UK). Mouse antibody against total moesin and the FLAG epitope were from Cell Signaling Technology (Beverly, MA, USA). Antibody targeting NG2-Cy3 conjugate was purchased from Millipore (St. Louis, MO, USA). The following antibodies were from Santa Cruz (CA, USA): rabbit anti-PDGFR-β, mouse anti-GFAP and mouse anti-von Willebrand factor (vWF).
Secondary antibodies for immunoblotting were manufactured by Sigma (St. Louis, MO, USA). FITC-anti-rabbit IgG second antibody was from Molecular Probes (Life Technologies, Carlsbad, CA, USA) and mouse anti-α-SMA was from Sigma. ROCK inhibitor Y27632 was from TargetMol (USA). The Cell Counting Kit (CCK)-8 was from Dojindo Laboratories (Kumamoto, Kyushu, Japan). Other chemicals were from Sigma unless otherwise indicated.

After the exposure, intestinal organoids were lysed in 100 μL RIPA buffer and the extracted proteins were denatured using SDS loading buffer (125 mM Tris–HCl pH 6.8, 4% sodium dodecyl sulfate, 20% glycerol, 0.04% bromophenol blue, and 100 mM β-mercaptoethanol) at 95 °C for 5 min. For each sample, 10 μg protein was loaded and separated on 10% or 12% mini-protein TGX precast protein gels (Bio-Rad) and transferred to polyvinylidene difluoride membranes (GE Healthcare, Chicago, USA). The membranes were incubated overnight at 4 °C with appropriate primary antibodies, subsequently with specific secondary antibodies and visualized using the enhanced chemiluminescence reagent (Thermo Scientific). The following antibodies were used: Rabbit anti-alpha-TUBULIN, mouse anti-E-cadherin, and rabbit anti-NF-κB p65 were from Abcam; rabbit antibodies against p38, phospho-p38, JNK, phospho-JNK, ERK, phospho-ERK, MLC, phosphor-MLC, phospho-NF-κB p65, STAT1, and phospho-STAT1 (Tyr701) were from Cell Signalling Technology; mouse anti-CLDN-2 and rabbit anti-ILDR-1 were from Invitrogen; anti-mouse/rabbit IgG horseradish peroxidase-linked secondary antibodies were from Cell Signalling Technology. Original western blot images were included in Supplementary Figures 7, 8, and 9. Densitometric quantification analyses of the western blots were performed using the Image J software.