Effects of Individual Micronutrients on CACO-2 Tight Junctional Proteins.

<p>Summary of effects of various micronutrients on a panel of eight tight junctional proteins in CACO-2 cell layers. Post-confluent CACO-2 cell layers in Falcon 75 cm² culture flasks were refed on both sides with control medium or medium containing 100μM Zinc 48 hrs before harvesting in lysis buffer. These total cell lysates were analyzed by PAGE followed by immunoblotting as described in Methods. Immunoblots were probed with primary antisera against specific tight junctional antigens also as described. Densitometry was performed on developed blots to quantitate band densities. Data shown represents the mean ± standard error for an n = 3 cell layers in all cases. P values are listed for instances of statistical significance (Student’s t test [two tailed] to at least the P<0.05 level). NS indicates non significance. This same procedure was used for quercetin (400μM), butyrate (5.0mM), berberine (100μM), and indole (1.0mM) at the concentrations that provided maximal barrier enhancement and for the same time periods used in Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.g001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.g002" target="_blank">2</a>. Significant increases are highlighted in green while significant decreases are highlighted in yellow to emphasize the distinct overall pattern specific to each micronutrient.</p><p>Effects of Individual Micronutrients on CACO-2 Tight Junctional Proteins.