The TEER assay was performed as described previously.38 Caco-2/BBE cells were cultured on 24-well 12 mm polyester Transwells (Corning, Corning, NY) for 21 days. IL-6 (10 ng/ml) was added to both the upper and lower chambers of the transwell cultures. TEER was measured before and after treatment for 24 hours ± UNC0638 (500 nM) or BRD4770 (10 μM) or vehicle using an EVOM2 epithelial voltage meter (World Precision Instruments, Sarasota, FL). TEER values were calculated after subtraction of the intrinsic resistance of the cell-free filter. To measure dextran permeability, 4 kDa fluoroscein isothiocyanate (FITC)-dextran (FD4; 3 mg/ml) was added to the upper chamber without medium change. Aliquots were withdrawn from the lower chambers after 4 h and assayed for fluorescence at 515 nm with excitation at 492 nm.
Evom2 epithelial voltage meter
Manufactured by World Precision Instruments
The EVOM2 epithelial voltage meter is a laboratory instrument designed to measure the transepithelial electrical resistance (TEER) of cell monolayers. It provides a reliable and accurate way to assess the integrity and barrier function of epithelial cell layers in cell culture systems.
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2 protocols using evom2 epithelial voltage meter
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Measuring Epithelial Barrier Function
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Transepithelial Electrical Resistance Measurement
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TEER values were measured with a STX2/chopstick electrode across the epithelial layer in Transwell inserts using an EVOM2 epithelial voltage meter (World Precision Instruments). According to the manufacturer’s instructions, electrodes were sterilized and adjusted before the measurement. To eliminate excessive mucus, cells were manually washed once with pre-warmed DPBS prior to TEER measurements. Subsequently, pre-equilibrated ALI medium (Stemcell Technologies) was added to the apical (0.25 ml) and basolateral (1 ml) chambers. Before reading TEER values, the monolayers were given time to reach a stable potential for ∼5 min inside the incubator. The sample resistance was calculated by subtracting the values from the cell-free Transwell insert’s resistance. Resistance of each Transwell insert in units of Ω was calculated using a previously described technique [38 (link)].
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