Fig. 5

pNKCC1 expression is increased in the CP of Ptpn20^−/− mice. A Expressions of Ptpn20, water and ion transporters in the CP of Ptpn20^−/− mice as analyzed by real-time PCR. Relative fold change is calculated using the 2^−ΔΔCt method. Data are normalized to β-actin and fold change is expressed relative to the expression level of Ptpn20 in the CP of WT mice set as 1. We did not find significant differences in expressions of NKCC1, AQP1 or Na⁺/K⁺-ATPase in CP between 8-week-old WT and Ptpn20^−/− mice. Values are as follows: NKCC1 = 1.18; AQP1 = 1.01; ATP1a1 (Na⁺/K⁺-ATPase) = 0.83 (n = 5 mice per group). B Immunoblotting of pNKCC1 protein expression in the CP of 8-week-old WT and Ptpn20^−/− mouse. Expression of pNKCC1 is higher in the CP of Ptpn20^−/− mice than in WT mice. Actin is used as the loading control (n = 5 mice per group). C Immunofluorescence of Ptpn20^−/− mouse CP illustrating expression of pNKCC1 in the membrane facing the lumen (green) with E-cadherin (red) as a basolateral marker. Scale bar = 20 μm. D Immunofluorescence of pNKCC1, NKCC1, AQP1 and Na⁺/K⁺-ATPase in 4-week-old WT and Ptpn20^−/− mice in the CP of the lateral ventricles and fourth ventricle are immunolabeled as shown. Ptpn20^−/− mice show stronger pNKCC1 expression in the cytoplasm and apical surface of CP epithelial cells than WT mice. No significant differences are seen in NKCC1, AQP1 or Na⁺/K⁺-ATPase between WT and Ptpn20^−/− mice. Scale bar = 10 μm