JCI - Inactivation of Notch signaling in the renal collecting duct causes nephrogenic diabetes insipidus in mice

Inactivation of Notch signaling in the renal collecting duct causes nephrogenic diabetes insipidus in mice

Hyun-Woo Jeong, Un Sil Jeon, Bon-Kyoung Koo, Wan-Young Kim, Sun-Kyoung Im, Juhee Shin, Yunje Cho, Jin Kim, Young-Yun Kong

Published in Volume 119, Issue 11

J Clin Invest. 2009; 119(11):3290–3300 doi:10.1172/JCI38416

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Figure 1

Inactivation of Notch signaling in the collecting duct of Hoxb7-CreMib1^f/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 Mib1^f/f (D) and Hoxb7-CreMib1^f/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 Mib1^f/f and Hoxb7-CreMib1^f/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 Mib1^f/f and Hoxb7-CreMib1^f/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 Mib1^f/f and Hoxb7-CreMib1^f/f littermates. Error bars represent mean ± SD. *P < 0.05.