The human tight junction protein 1 (TJP1) ELISA kit (MyBiosource, San Diego, CA, USA) was used following the manufacturer’s instructions [53 (link)]. CaCo-2 cells were lysed with cold PBS (Merck Life Science, Rome, Italy) 1×, centrifuged at 5000× g for 5 min at 4 °C, and 100 μL of each sample were analyzed. The plates were read by a spectrometer (Infinite 200 Pro MPlex, Tecan, Männedorf, Switzerland) at 450 nm. The data were obtained and compared to the standard curve (ranging from 0 to 1000 pg/mL), and the results were expressed as a mean ± SD (%) versus the control (0 line) of five independent experiments performed in triplicate.
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