Acrylamide bis acrylamide solution

Nutrient broth and chemicals: Coomassie brilliant blue (CBB) R-250, 2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris), Bradford reagent, bovine serum albumin (BSA) and acrylamide/bis-acrylamide solution were purchased from HiMedia (Mumbai, India). Low-molecular-mass marker was purchased from GeneDireX (Taoyuan, Taiwan). N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Suc-AAPF-pNA), azocasein, gelatin, 4-aminobenzamidine dihydrochloride, dithiothreitol, sodium dodecyl sulfate (SDS), glycerol, bromophenol blue, trichloroacetic acid (TCA), ethylenediaminetetraacetic acid (EDTA), Triton X-100, ammonium sulfate, sodium acetate, sodium phosphate, glycine, NaCl, NaOH, HCl, KCl, CaCl₂, CuCl₂, ZnCl₂ and MgCl₂ were purchased from Sigma-Aldrich, Merck (St. Louis, MO, USA).

DMEM high glucose, fetal bovine serum (FBS), penicillin-streptomycin, trypsin-EDTA, L-glutamine, and Transwell inserts (0.64 mm diameter, 0.4 µm pore size, Costar) were purchased from Euroclone (Milan, Italy). Minimum essential medium non-essential amino acids (MEM-NEAA), SuperSignal^TM West Dura Extended Duration Substrate, Pierce^® IP lysis buffer, Pierce^TM Coomassie Plus Assay Reagent, LPS from E. coli 026:B6, TRIzol^®, High-Capacity Reverse Transcription Kit, and PowerUp^TM SYBR^TM Green Master Mix were obtained from Thermo Fisher Scientific (Rockford, IL, USA). Skim milk powder was purchased from Microgem (Pozzuoli, Italy). Acrylamide/bis-acrylamide solution 30% (29:1) was obtained from HiMedia Laboratories (Nashik, India). Primary monoclonal antibodies against CDH1 (4A2), CLDN-1 (D5H1D), CLDN-2 (E1H90), OCLN (E6B4R), and HRP-linked anti-mouse and anti-rabbit IgG were obtained from Cell Signaling Technology (Waltham, MA, USA). The primary antibody against β-Tubulin (E-AB-20095) was purchased from Elabscience (Huston, TX, USA).