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Fig. 4. Phosphorylation of AQP2 by DDAVP in CnA
+/+ and -/- mice. (A) P-AQP2 was detected by immunoblotting with a specific antibody in medullary protein lysates from +/+ mice treated either with vehicle control or DDAVP for 1 hour. Actin was included as a control. Lanes 1-3, untreated; lanes 4-7, plus DDAVP. Results from typical immunoblots were evaluated by semi-quantitative analysis and are shown in bar graph as the mean ± s.e.m. *P<0.05 (Student's t-test). (B) P-AQP2 was detected in medullary protein lysates from -/- mice treated with either control or plus DDAVP for 1 hour. Actin was included as an internal control. Lanes 1-3: control mice; and lanes 4-7: plus DDAVP. Results from typical immunoblots were evaluated by semi-quantitative analysis and results are shown as bar graphs as the mean ± s.e.m. (C) Expression of PKA was determined by western blotting using a specific antibody in +/+, +/- and -/- mouse kidney-cell homogenates. Results from two independent experiments were evaluated by semi-quantitative analysis and are shown as bar graphs as the mean ± s.e.m. (D) In an in vitro assay (see Materials and Methods) activity of PKA was determined in homogenates of whole kidneys of CnA +/+ and -/- mice that had been either treated with vehicle only (control) or with DDAVP for 1 hour. Bars show the mean ± s.e.m. of 4-6 animals per genotype. *P<0.05 compared with +/+ control (Student's t-test). (E) Intracellular localization of AQP2 was examined by immunohistochemistry in the inner strip of the outer medulla in both +/+ and -/- mice following DDAVP treatment. Arrows identify AQP2 apical localization in +/+ and cellular/baso-lateral distribution in -/- mice. Magnification is 40 x. Results of 4-6 mice per group were evaluated by semi-quantitative analysis and are shown in bar graph as the mean ± s.e.m. *P<0.05.
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