Figure 1
Inactivation of Notch signaling in the collecting duct of
Hoxb7-CreMib1f/f mouse kidneys.
(A and B) Immunohistochemical staining of E16.5 C57BL/6 mouse embryonic kidneys with anti-Jag1 (A, green), anti-Notch2 (B, green), and anti-AQP2 (A and B, red) antibodies. Hoechst (blue) stains DNA, and dotted lines indicate renal collecting ducts. Note the different fluorescent intensities of Jag1 in the collecting duct (A, arrowheads). Scale bars: 15 μm. (C) Immunohistochemical staining of P0 TNR mouse kidneys with anti-EGFP (green) and anti-AQP2 (red) antibodies. In the collecting ducts, EGFP signals are not detected in AQP2-negative cells (arrowheads). Hoechst (blue) stains DNA, and dotted lines indicate renal collecting ducts. Scale bar: 15 μm. (D and E) Immunohistochemical staining with anti-Mib1 (green) and anti-AQP2 (red) antibodies in the kidneys of E18.5 Mib1f/f (D) and Hoxb7-CreMib1f/f (E) littermates. Hoechst (blue) stains DNA, and dotted lines indicate renal collecting ducts. Scale bars: 30 μm. (F) Quantitative real-time RT-PCR analysis of Mib1 in the kidneys of E18.5 Mib1f/f and Hoxb7-CreMib1f/f littermates. Error bars represent mean ± SD. *P < 0.05. (G) Western blot analysis of Notch2 intracellular domain (N2ICD) in the kidneys of E18.5 Mib1f/f and Hoxb7-CreMib1f/f littermates. Numbers indicate relative values of intensity normalized to β-actin. The arrowhead indicates the 77-kDa protein. (H) Quantitative real-time RT-PCR analysis of HeyL in the kidneys of E18.5 Mib1f/f and Hoxb7-CreMib1f/f littermates. Error bars represent mean ± SD. *P < 0.05.
