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. 2011 Oct 14;6(10):e25847. doi: 10.1371/journal.pone.0025847

Figure 4. e15.5 GSK-3β null embryos exhibit increased canonical Wnt signaling in the developing palate.

Figure 4

(A) Immunohistochemistry for total β-catenin (second column), active β-catenin (third column), and Axin-2 (fourth column) in GSK-3β +/+ and GSK-3β −/− embryos. The first column is included for orientation purposes. The dotted lines in the first column indicate the areas shown in higher magnification in the second, third, and fourth columns. As shown, GSK-3β −/− embryos have more intense total β-catenin, active β-catenin, and Axin-2 immunostaining than controls in the palatine bone (pb), suggesting that GSK-3β −/− embryos have increased canonical Wnt signaling compared to GSK-3β +/+ embryos. Tongue (t) and nasal cavity (nc) are labeled for orientation purposes. Scale bars in the lower magnification images (first column) represent 200 µm. Scale bars in the higher magnification images (second, third, and fourth columns) represent 100 µm. (B) Quantification of DAB-positive nuclei per high power field, displaying significantly greater active β-catenin positive nuclei in the GSK-3β −/− palates when compared to controls. N = 3, *<p0.05. (C) In situ hybridization for Wnt 9b, a canonical Wnt ligand expressed in the craniofacial region. The dotted lines in the first and fourth column indicate the area shown in higher magnification in the second and third columns, respectively. The intensity and distribution of Wnt9b transcripts is greater in the GSK-3β −/− palates when compared to controls. Tongue (t) and nasal cavity (nc) are labeled for orientation purposes. Scale bars in the lower magnification images (first and fourth column) represent 200 µm. Scale bars in the higher magnification images (second and third columns) represent 100 µm.