Fig. 8.

There was decrease in the beclin 1/Bcl-2 complex in the kidneys of Tg-Kl mice. Tg-Kl mice and their wild-type (WT) littermates; Becn1^+/− and chimeric mice with Tg-Kl (Becn1^+/−;Tg-Kl) as well as Bcl2^AAA mice and chimeric mice with Tg-Kl (Bcl2^AAA;Tg-Kl) were fasted for 24 h. At the final 4 h, mice were intraperitoneally injected with chloroquine once. Twenty-four hours after starvation initiation, mice were euthanized, and kidneys were harvested for experiments. A: beclin 1/Bcl-2 complex and autophagic status in the kidney. Top, immunoprecipitation (IP) with Bcl-2 antibody followed by immunoblots (IBs) for beclin 1 and Bcl-2. Bottom, representative immunoblots for Klotho, beclin 1, Bcl-2, light chain (LC)3, and p62 in total input proteins. B: summary of the changes in markers from all immunoblots. C: green fluorescent protein (GFP)-LC3 puncta in the kidneys by immunohistochemistry. Top, representative microscopic images of LC3 puncta. Scale bar = 50 μm. Bottom, quantitative analysis of the number of GFP-LC3 punctas in renal tubules. Data are expressed as means ± SD, and statistical significance was evaluated by two-way ANOVA followed by a Student-Newman-Keuls post hoc test and significance was accepted when *P < 0.05 and **P < 0.01 between two groups. Sample numbers in each group are presented in brackets underneath the corresponding bars.