Figure 3. Wnt3aCM but not Wnt5aCM... | Rockefeller University Press

$Figure 3. Wnt3aCM but not Wnt5aCM induced TJs and barrier properties in MBEs. Wild-type MBEs treated with controlCM, Wnt3aCM, or Wnt5aCM in vitro. (A, a, e, and i) IHC staining for Cldn3. (b, f, and j) IHC staining for Cldn5. Panels are the same magnification as indicated in panel k. (c, g, and k) IHC staining for active β-cat (clone 8E7). Arrowheads point to positive nuclei. (d, h, and l) Freeze-fracture EM. Arrowheads point to P-face–associated TJ ridges. (B) mRNA levels for Cldn1, Cldn3, Cldn5, Cldn12, Ocln, Plvap, and Axin2 measured by quantitative RT-PCR. Cldn3 (n = 3; *, P = 0.0319), Plvap (n = 3; *, P = 0.0098), and Axin2 (n = 3; **, P = 0.005); other tested genes were not significantly altered. Control is set to 100% (dashed line). (C) Western blotting for Cldn3 and Cldn5. α-Tubulin/tubulin α1A served as a loading control. Western blotting for 8E7 and total β-cat. No band could be detected in Wnt5aCM-treated cells. See graphs for densiomeric quantification of bands. Error bars represent SEM. Data are representative of at least three independent experiments.$

Figure 3.

Wnt3aCM but not Wnt5aCM induced TJs and barrier properties in MBEs. Wild-type MBEs treated with controlCM, Wnt3aCM, or Wnt5aCM in vitro. (A, a, e, and i) IHC staining for Cldn3. (b, f, and j) IHC staining for Cldn5. Panels are the same magnification as indicated in panel k. (c, g, and k) IHC staining for active β-cat (clone 8E7). Arrowheads point to positive nuclei. (d, h, and l) Freeze-fracture EM. Arrowheads point to P-face–associated TJ ridges. (B) mRNA levels for Cldn1, Cldn3, Cldn5, Cldn12, Ocln, Plvap, and Axin2 measured by quantitative RT-PCR. Cldn3 (n = 3; *, P = 0.0319), Plvap (n = 3; *, P = 0.0098), and Axin2 (n = 3; **, P = 0.005); other tested genes were not significantly altered. Control is set to 100% (dashed line). (C) Western blotting for Cldn3 and Cldn5. α-Tubulin/tubulin α1A served as a loading control. Western blotting for 8E7 and total β-cat. No band could be detected in Wnt5aCM-treated cells. See graphs for densiomeric quantification of bands. Error bars represent SEM. Data are representative of at least three independent experiments.

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