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. 2022 Jul 21;119(30):e2202125119. doi: 10.1073/pnas.2202125119

Fig. 3.

Fig. 3.

Lrba knockout mice display the NDI phenotype. (A) LRBA interacts with PKA RIIα and RIIβ in vitro. FLAG-tagged PKA regulatory subunits and Myc-tagged LRBA were overexpressed in HEK293T cells. Anti-FLAG beads were used to perform coimmunoprecipitations. (B) LRBA interacts with PKA RIIβ in vivo. Membrane fraction of mouse kidneys was prepared. Anti-RIIβ antibody was used to perform a coimmunoprecipitation assay. (C) The baseline urine-concentrating ability is impaired in Lrba−/− mice. Metabolic cages were used to monitor urine osmolality, urine volume, and water intake for 24 h. (n = 5 WT mice; n = 4, He mice; n = 7 Ho mice). (D and E) The maximal urine-concentrating ability is impaired in Lrba−/− mice. Mice were deprived of water for 12 h. (D) Left, urine osmolality at the beginning and end of the experiments (n = 6). Right, representative urine samples. (E) Percentage of weight loss after water deprivation test (n = 6). (F) Urine osmolality of Lrba−/− mice is unresponsive to vasopressin. Left, changes in urine osmolality of WT and Lrba−/− mice after the treatment of dDAVP (0.4 μg/kg) (n = 7). Right, representative urine samples. (C–E) Data are reported as mean ± SD. **P < 0.01, NS, not significant by Dunnett's test. (F) **P < 0.01 by two-sided Student's t test. Ab, antibody; He, Lrba+/−; Ho, Lrba−/−; IP, immunoprecipitation.