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. Author manuscript; available in PMC: 2010 May 1.
Published in final edited form as: Mech Dev. 2009 Mar 4;126(5-6):337–349. doi: 10.1016/j.mod.2009.02.005

Figure 7. NRAGE modulated both the Ret and BMP signaling in the developing kidney.

Figure 7

(A) Ret protein expression increases as NRAGE expression decreases. Lysates were generated from three E11.5 kidney explants per well cultured with 10 μM of NRAGE morpholino for three days and analyzed via western blot analysis. Antibodies to Ret, NRAGE, and β-actin were utilized. (B) NRAGE interacts with Ret receptor in mIMCD-3 receptors. Lysates were generated from mIMCD-3 cells treated with and without 25 ng/ml of GDNF for 1 hour. Immunoprecipitation with NRAGE and subsequent western blot analysis demonstrates that NRAGE interacts with Ret with and without GNDF addition. (C-G) Addition of GDNF to NRAGE morpholino treated kidneys stimulates branching of the UB. GDNF was added to NRAGE morpholino (D) or negative control morpholino (F) treated E11.5 ICR kidney explants for three days. Extent of the effect of GDNF addition on morpholino treated kidneys was determined by anti-laminin staining and counting branch end points of each explants. (G) There was a significant increase in the number of branch endpoints in kidney explants treated with NRAGE morpholino and GDNF (*unpaired two-tailed t-test p=0.011). Presented are the means of six explants per treatment.