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. 2019 Jun 1;30(12):1406–1424. doi: 10.1091/mbc.E19-01-0008

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© 2019 Van Itallie et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 9: — Cldn2 relocalizes to cell contacts after calcium switch assay more slowly than does cldn1 or cldn4. (A) MDCK II cells cultured on semipermeable membranes were incubated in low-calcium media for 15 h; calcium was added back, and cells were collected and fixed for immunofluorescence at various times after readdition. TER was measured in parallel. (A) Immunofluorescence analysis—(left panel) cldn2, cldn1, and ZO-1 and (right panel) cldn4, ocln, and ZO-1 in untreated monolayers (top images), after 15 h in low calcium (second row), 6 h after calcium readdition (third row), and 24 h after calcium readdition—reveals that by 6 h of calcium return, most the cldn1, cldn4, ocln, and ZO-1 but not cldn2 has returned to cell contacts; the difference in cell-contact localization between cldn2 and 4 is quantified in B. (C) TER measurements show that measurable TER is lost in low calcium but overshoots control levels by more than 10-fold before returning to control values at 24 h.