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Figure 6


Fig. 6. Expression of total and p-AQP2 in IMCD-cell vesicle and membrane fractions. (A) IMCD-cell vesicle fractions (VFs) were isolated from +/+, +/- and -/- mice. Expression of total and p-AQP2 was detected by immunoblotting with specific antibodies. Actin was included as a control. Lanes 1-3, +/+; 4-6, +/-; and 7-9, -/-. Levels of total and p-AQP2 were evaluated by semi-quantitative analysis, normalized against actin and shown in bar graph. Data show the mean ± s.e.m. of data from 3-5 mice. *P<0.05 (Student's t-test). (B) Calcineurin activity and expression were examined by western blotting VFs with an antibody that crossreacts with the phosphatase domain of all three A subunit isoforms. Actin expression was examined in the same samples as an internal control. (C) Calcineurin activity in isolated VFs was determined in an vitro assay. Bar graphs show the mean ± s.e.m. of 4-6 samples per group. *P<0.01 (Student's t-test). (D) MFs were prepared and expression of total and p-AQP2 were found by direct immunoblotting. Lanes 1-3, +/+; lanes 4-6, +/-; lanes 7-9, -/-. Actin was included as a control. Levels of total and p-AQP2 were evaluated by semi-quantitative analysis and results are shown in bar graphs as the mean ± s.e.m. of data from 3-5 mice. *P<0.05, **P<0.01 (Student's t-test).





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