Victor x2 2030 fluorimeter

IPEC-J2 cells were seeded on 6-well, 0.4 µm pore-size polyester membrane inserts and were grown to confluent, differentiated monolayers. Transepithelial electrical resistance (TEER) measurement of monolayers was performed on alternate days after seeding, from day 7 of culture, using an EVOM Epithelial Tissue Volt/Ohmmeter (World Precision Instruments, Berlin, Germany) [28 (link),29 ]. S. Typhimurium LPS was added at 10 µg/mL concentration alone and with combinations of Q, QM, and R in two concentrations (25 µM and 50 µM). At the same time as LPS and combination treatments administration, 1 mg/mL fluorescein isothiocyanate dextran 4 kDa (FD4) tracer dye was added to the cells (Sigma-Aldrich, Darmstadt, Germany) with different incubation times (2 and 4 h). Supernatants from the basolateral chambers were collected, and the FD4 concentration was measured by a fluorescent method at excitation 485 nm and emission 535 nm (Perkin Elmer, Victor X2 2030 fluorimeter).

IPEC-J2 cells were seeded on 6-well polyester membrane inserts and were grown to confluent, differentiated monolayers. LPS derived from S. Typhimurium was added at 10 μg/ml concentration for 1 h to cells, and transepithelial electric resistance (TEER) values were measured prior to and 2, 4, and 24 h after LPS treatment. Simultaneously with LPS administration, cells were treated with 1 mg/ml fluorescein isothiocyanate-dextran 4 kDa (FD4) tracer dye (Sigma-Aldrich, Darmstadt, Germany) with different incubation times (2, 4, and 24 h). Medium samples from the basolateral chambers were collected, and the FD4 concentration was determined by a fluorescent method at excitation 485 nm and emission 535 nm (PerkinElmer, Victor X2 2030 fluorimeter).