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. Author manuscript; available in PMC: 2010 Jul 2.
Published in final edited form as: Nat Med. 2009 Aug 30;15(9):1046–1054. doi: 10.1038/nm.2010

Figure 2.

Figure 2

Jbn is required for Wnt activity in adult mouse kidney. (a) X-gal staining in 5-month-old littermate kidneys. Cross-section reveals outer stripe (os) staining. OM, outer medulla; CO, cortex; is, inner stripe. Dashed line, calyx boundary. (b) Top, antibody staining for β-galactosidase (green) in 5-month-old Ahi1+/− TOPGAL+ or Ahi1−/− TOPGAL+ kidneys. Hoechst labels nuclei (blue). Bottom, β-galactosidase average fluorescence. *P < 0.05, Student's t test, n = 3 images. Error bars represent means ± s.e.m. (c) β-galactosidase (green) and Lef-1 (red) protein staining (arrows) in Ahi1+/− TOPGAL+ and Ahi1−/− TOPGAL+ mice. Inset provides higher magnification of Lef-1 staining. Arrowheads indicate negative tubules for reference. (d) Axin-2 and DKK-1 staining (red, arrows) in the corticomedullary region of littermate kidneys. (e) Top, Lef-1 western blot from whole kidney lysates of littermates at 5 months and 1 year of age, revealing decreased expression of the full-length isoform (55–57 kDa, arrow). Bottom, full-length Lef-1 measurement relative to α-tubulin (loading control). (f) X-gal and β-galactosidase antibody staining in littermate kidneys at 2.5 months of age, before the onset of pathology (H&E at right). Dashed line demarcates the medullary boundary. (g) Lef-1 target gene staining (green) of Ahi1+/− and Ahi1−/− mice before nephronophthisis pathology (H&E at left). (h) Top, Lef-1 and GAD-1 western blots from whole-kidney lysates of Ahi1+/+ and Ahi1−/− littermates at 3 months of age. Bottom, full-length Lef-1 and GAD-1 levels relative to α-tubulin.