(A) Kaplan-Meier survival curves of different genotyping mice. WT (n=52), Bk/BK (n=61), BK/BK;kl/kl (n=63), and kl/kl (n=43) mice. *P<0.05 vs BK/BK, ##P<0.01 vs kl/kl mice by log-rank (Mantel-Cox) test. (B) Comparison of body weight of WT, Bk/BK, BK/BK;kl/kl, and kl/kl genotypes between male and female mice at the age of 10 weeks. The sample size of each genotype is shown in parentheses. Data are presented as mean ± S.D. with scatter plots of individual data points. **P<0.01 vs kl/kl mice by one-way ANOVA followed by Student-Newman-Keuls post hoc test. (C) Change in plasma phosphate (Pi) in WT (n=46 in total including: male 23; female: 23); Bk/BK (n=40 in total including male: 20; female: 20); BK/BK;kl/kl (n=38 in total including male: 18; female 20); and kl/kl mice (n=28 in total including male: 13; female: 15). *P<0.05, **P<0.01 vs BK/BK mice, #P<0.05, ##P< 0.01 kl/kl mice groups by one-way ANOVA followed by Student-Newman-Keuls post hoc test at the indicated time point. (D) Immunoblots of total kidney lysates for LC3 and p62 protein in the kidneys of 4 genotypes at the age of 6 and 10 weeks, respectively. Upper panel shows representative immunoblots. Bottom panel summarizes data presented as mean ± S.D. Each group has 5 mice at each time point. *P<0.05, **P<0.01 between two groups by one-way ANOVA followed by Student-Newman-Keuls post hoc test. (E) Change in plasma αKlotho in WT; Bk/BK; BK/BK;kl/kl; and kl/kl mice at the age of 4, 6, 8 and 10 weeks respectively. The data were presented as mean ± S.D. with scatter plots of individual data points. Each genotype has 5 mice at each time point. *P<0.05 vs BK/BK and ##P<0.01 vs kl/kl by one-way ANOVA followed by Student-Newman-Keuls post hoc test at 10 weeks-old. (F) Renal αKlotho expression. Upper panel shows representative immunoblots of total kidney lysates for αKlotho protein in the kidney of 4 genotypes (kl/kl, WT, BK/BK;kl/kl, and BK/BK mice) at the age of 4, 6, 8 and 10 weeks respectively. Each genotype has 5 mice at each time point. Bottom panel is a summary of all blots and presented as mean ± S.D. with scatter plots of individual data points. *P<0.05, **P<0.01 between 2 groups at same time point by one-way ANOVA followed by Student-Newman-Keuls post hoc test.