Sc 51322

CaCl₂ was purchased from Bodi Chemical Co., Ltd. (Tianjin, China). Antibodies specific against NeuN and Alexa Fluor-488, Alexa Fluor-555, and HRP-labeled secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, United States). S(+)-Ketamine (60 mg/kg, for 1 h), SC-51322 (30 nM, for 12 h), PF-04418948 (100 nM, for 12 h), and DG-041 (60 nM, for 12 h) were obtained from R&D Systems (Minneapolis, MN, United States), and CJ-42794 (40 nM, for 12 h) was obtained from MedChem Express (Monmouth Junction, NJ, United States). High-fidelity (HF) restriction enzymes for EcoRI, BamHI, XhoI, and AgeI were purchased from New England Biolabs (Beverly, MA, United States). DAPI was procured from Beyotime Institute of Biotechnology (Haimen, China). The plko.1-puro, psPAX₂, pMD₂.G, and plvx-IRES-zsgreen vectors were purchased from Addgene (Sidney, SD, United States). All the reagents used for the quantitative (q)RT-PCR and SDS-PAGE experiments were purchased from Bio-Rad Laboratories (Hercules, CA, United States), and all other reagents were obtained from Invitrogen (Carlsbad, CA, United States), unless otherwise specified.

Intestinal microvascular endothelial cells (MIMECs) were isolated and cultured as reported previously^{4 (link),5 (link)}. When the MIMECs were cultured for 7 days, we obtained almost pure cultures of MIMECs (90% confluence). The MIMECs were subjected to the preliminary dosage selection assay, and the current dosage of TNF-α (30 ng/mL) was selected for 12 h for subsequent experiments. For pharmacological PGE2 management and inhibition of various PGE2 receptors, the culture system was further cultured with the following reagents and final concentrations: stabilized PGE2 analog 16,16-dimethyl PGE2 (dmPGE2, 1 μM, R&D Systems), EP1 inhibitor (10 μM, SC 51322; R&D Systems), EP2 inhibitor (10 μM, PF 04418948; R&D Systems), EP3 inhibitor (10 μM, L-798,106; R&D Systems), and EP4 inhibitor (10 μM, L-161,982, R&D Systems).