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J Cell Sci. Author manuscript; available in PMC 2012 Jun 6.
Published in final edited form as:
J Cell Sci. 2004 Feb 1; 117(Pt 4): 559–570.
Published online 2004 Jan 6. doi: 10.1242/jcs.00893
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Fig. 4

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Fractionation of MDCK cells in iodixanol gradients. MDCK cells were homogenized either 6 hours or 48 hours after induction of calcium-dependent cell-cell adhesion. Post-nuclear supernatants were mixed with 10%, 20% and 30% (w/v) iodixanol, layered step-wise in centrifuge tubes and centrifuged at 350,000 g for 3 hours. The presence of Sec8, ZO-1, ZO-2, afadin, occludin, claudin-1, claudin-2, E-cadherin, nectin-1α and nectin-2α in gradient fractions was assayed by SDS-PAGE followed by immunoblotting with specific antibodies. Protein levels were quantified using a Molecular Dynamics Phosphorimager. Densities of each fraction were calculated after measuring refractive indices with a refractometer, and are plotted as dotted lines on each graph with values (in g/ml) indicated on the y-axis.

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