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21-BD regulates tight junctions.

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posted on 07.10.2014 by Sayonarah C. Rocha, Marco T. C. Pessoa, Luiza D. R. Neves, Silmara L. G. Alves, Luciana M. Silva, Herica L. Santos, Soraya M. F. Oliveira, Alex G. Taranto, Moacyr Comar, Isabella V. Gomes, Fabio V. Santos, Natasha Paixão, Luis E. M. Quintas, François Noël, Antonio F. Pereira, Ana C. S. C. Tessis, Natalia L. S. Gomes, Otacilio C. Moreira, Ruth Rincon-Heredia, Fernando P. Varotti, Gustavo Blanco, Jose A. F. P. Villar, Rubén G. Contreras, Leandro A. Barbosa

MDCK cells were cultured in transwell permeable supports and treated with 5, 10 and 50 µM 21-BD. (A) TER was measured as a function of time. The control TER data (white circles, dotted line) averaged 183±8 Ω.cm2 (n = 13) and were normalized to 100%. 5 and 10 µM 21-BD provoke transient small increases of TER, while 50 µM 21-BD causes a stronger and a sustained TER increase (red circles). (B) MDCK cells were incubated 48 h with different concentrations of 21-BD (red symbols) or digoxin (green symbols). mRNA cell content of claudins -4 (circles) and -2 (triangles) were measured by quantitative real time PCR. (C) Protein cell content of the tight junction integral membrane proteins claudins -4 and -2 and the membrane-associated protein ZO-1 as a function of 21-BD concentration in the media for 48 h. Images from the left part of the figure C are representative immunoblots and the graph in the right part is the statistical analysis.