Figure 6. Immunohistochemical analysis on Ret phosphorylation.

(A–E) Immunohistochemical analysis of lower urinary tracts with anti–Ret (pY1015) (green) (A and B), anti-Ret (green) (D and E), anti–E-cadherin (red) (A, B, D, and E), and no primary antibodies (C). Nuclei are stained with DAPI (blue). Green channel images from white line squares are presented in the upper right corners. (F) Quantification of immunofluorescent signals in E12.5 CND. Ret (pY1015) was significantly hyperphosphorylated in Ptprs^–/–Ptprf^ΔP/ΔP CNDs (B and F) in comparison with control CNDs (A). Note that there were no significant differences in Ret or E-cadherin expression levels between control and Ptprs^–/–Ptprf^ΔP/ΔP CND (F). Dotted lines outline the cloaca epithelium. (G and H) In situ hybridization analysis of E12.5 transversal sections with antisense Ret cRNA probes. Ret expression was present in the nephric duct but strongly downregulated in both control (G) and Ptprs^–/–Ptprf^ΔP/ΔP (H) CNDs. Scale bars: 10 μm. The results shown are means ± SEM. Ab (–), no primary antibody.